Methods of DNA Identification

Methods of DNA Identification To isolate DNA from blood, saliva, buccal swab and betel quid by phenol-chloroform method and chelex method and compare the efficacy of both the methods. To carry out restriction digestion of the DNA samples isolated from above mentioned sources using the restriction enzyme EcoRI (G|AATTC) and identify individuals based on the pattern of restriction banding and to ascertain the applicability of the restriction digestion in forensics MATERIALS AND METHODS: Blood, saliva, betel quid and buccal swab were collected from 15 patients and DNA isolation was done by phenol-chloroform method and chelexmethod. DNA fingerprinting was carried out using EcoRI restriction enzyme. RESULTS:  DNA could be extracted from residues of saliva, DNA fingerprinting done with the isolated DNA was able to match with those of individuals. Chelex method was found to be more efficient than the Phenol-chloroform method KEY WORDS: Betel Quid, Chelex method, DNA,DNA fingerprinting,Phenol chloroform method Introduction DNA fingerprinting  has ascertained an increasingly imperative role towards decision making in judiciary. DNA tests have helped convict suspects, to exonerate suspects or overturned previous convictions. Scientific evidences such as fingerprints, blood, semen, shreds of clothing, hair, weapons, tire tracks, and other physical evidence at the crime scene can be a more riveting to a tribunal than the testimony of an eyewitness. DNA is more suitable because DNA remains scathe lessin challenging environments where such evidence is found. The DNA molecule holds an impressive dependability to withstand time. 1 DNA profiling compares the DNA fragment lengths and patterns. The isolated DNA from the samples is fragmented using a restriction enzyme. Then the length of the resulting fragments is determined by electrophoresis and comparedby a visual interpretation of the pattern of DNA bands. 2 DNA can be sourced from freshblood, fresh or dried human buccalswabs, soft tissue, saliva and salivary stains. Optimizing the methodology in DNA extraction from various sources have been tried by many studies. Minute quantities of saliva allows establishing DNAprofile. 3DNA has been proven to be isolated from cell samples from objects that was in contact with the body and from sources like chewing gums, cigarettes, bite marks in foods, among others. Restriction fragment length polymorphism (RFLP) analysis provides details of the DNA which is referred as a DNA fingerprint. As DNA is unique to every individual, analyzing the sequence helps in identification of specific patterns of each individual. DNA profile is considered as valid evidence in the court of law for paternity disputes and human identification. Standardization of DNA extraction technique will improve the reliability and speed up sample processing time. 4-6 Limited availability of biological samples in a crime scenechallenges the procedure of extraction , characterization and analysis of DNA. Furthermore, difficulty arises in retrieving DNA from stains and degraded samples which provide contaminated or poor qualityDNA. Hence, purification of DNA from samples is still a significant step in obtaining useful genotypes. Notwithstanding, tremendous advances have been made in the recent times in DNA testing. 7 Chewed betel quid (BQ) stains are encountered frequently on crime scenes in Southeast Asian countries. Though the quid presents as an important biological evidence, the forensic analysis using betel quid as an evidence has been impeded due to difficulty in extraction of human DNA . Hence, constituting a definite method for extracting DNA from chewed Betel quid residues is of paramount importance. 8 Saliva found on victims of several violent crimes is a potential source of DNA. They can be recovered from bite marks, cigarette butts, betel quid, postage stamps, envelopes and other objects. However , salivary stains usually dry up easily becoming invisible, making recognition and collection difficult. Among the various biological sources available, salivary analysis have great discriminatory power and can be incorporated into a criminal investigation . Improvisation of DNA extraction procedures will improve its reliability and also help to expedite the process. The present study aims to isolate DNA from blood, saliva (under different conditions) by phenol chloroform method and chelex method and to find the efficacy of these methods in extraction of DNA from traces of saliva. 9,10 ISOLATION OF DNAFROM BLOOD AND SALIVA BY PHENOL CHLOROFORM METHOD : The DNA was extracted with an equal volume of phenol: chloroform: isoamyl alcohol. This mixture was centrifuged at 10000rpm for 5 minutes. The aqueous phase was collected and extracted with chloroform: isoamyl alcohol mixture and centrifuged at 10000rpm for 5 minutes. The supernatant was transferred to a new microfuge tube and 0. 6 volume of isopropanol was added. The spongy white precipitate was transferred to a microfuge tube and added equal volume of ethanol was added. Then it was centrifuged at 10000rpm at room temperature for 10 minutes. The supernatant was drained and to the pellet 100 µL of TE buffer was added stored at 4 °C. ISOLATION OF DNAFROM BLOOD AND SALIVA BY CHELEX METHOD: 0. 5 ml of whole blood was collected in 2 ml tube and the cells are harvested by centrifugation at 3000 rpm for 3 min. at 4 °C. The supernatant was discarded. 0. 8 ml TBP buffer was added to the collection tube, vortexed gently, then centrifuged at 3000 rpm for 3 minutes, supernatant was discarded. The next stepwas continued if the blood pellet looks mauve 0. 5 ml of TBM buffer was added to the tube, and vortexed followed by addition of 3  µLof proteinase K and incubated at 55 °C for 30 minutes. The sample was centrifuged for 2 minutes at 5000 rpm and the supernatant saved to 2 ml tube and then added 260  µL of absolute ethanol. The mixture was applied to EZ-10 column, centrifuged at 8000 rpm for 1 minute; discarded the flow in the collection tube. 500  µL of wash solution was added and centrifuged at 8000 rpm for 1 minute. This step was repeated spin at 8000 rpm for an additional minute to remove residual amount of wash solution. The column was placed into a clean 1. 5 ml microfuge tube and 30  µL of elution buffer was added into the center part of membrane . The tube was incubated at 50 °C for 2 minutes centrifuged at 10,000 rpm for 1 minute to elute the DNA from the column The standards and samples were removed from the freezer and thawed. In a separate sterile 1. 5 ml microfuge tube for each standard/sample, 10  µl of DNA was mixed with 990  µl of D. I. water and vortexed . The solution was allowed to stand for 10 minutes to ensure the complete diffusion of DNA throughout the solution. This represents a 1:100 dilution of the standards and the DNA samples. B. DNAquantification The DNA sample was briefly vortexed and the solution wastransfered to the cuvette of the spectrophotometer with care not to create bubbles. The cuvette is inserted into the spec ensuring the correct face of the cuvette is in line the light beam. . An absorbance reading appears on the screen . Reading is continued until all standards and samples have been quantified. The concentration of DNA in the sample is determined according to the conversion factor (A260 of 1. 0 = 50  µg ml-1 DNA). The concentration of DNA in the sample can be read as  µg/mL using the conversion factor and dilution factor . RESTRICTION DIGESTION: Restriction enzyme buffer was vortexed before pipetting to ensure that it was well-mixed and was added to the tube . Appropriate amount of DNA to be cut wasvortexed before pipetting to ensure that it was well-mixed and was added to the tube. After vortexingthe enzyme to ensure that it was well-mixed 1 ÃŽ ¼L of enzyme EcoRIwas added. The mixture is placed in thermal cycler (Eppendorf) for2-3 hour incubation at 37 °C . To heat inactivate the enzyme the mixture is maintained at 80 °C for 20 min. The mixture is kept at 4 °C until the reaction mixture is out of the thermal cycler. Agarose Gel Electrophoresis Protocol Preparation of the agarose gel 1. 25 g Agarose powder was taken in 500 ml flask and 125 ml of TAE Buffer was added to it. The mixture is melted in hot water bath till a clear solution forms. The solution is allowed to cool to a temperature of 50-55 °C by periodic swirling to achieve even cooling. To it ethidium bromide solution was added. The ends of the casting tray are sealed with two layers of tape. The combs are placed in the gel casting tray. The melted agarose solution was poured into the casting tray and allowed to cool until it is solid. The comb and the tape are removed carefully. The gel is placed in the electrophoresis chamber. 2-3 mm of TAEBuffer is added over the gel. Loading the gel 6 à ¯Ã‚ Ã‚ ­l of 6X Sample Loading Buffer is added to each DNA sample containing tubes. 20 à ¯Ã‚ Ã‚ ­l of each sample is pipetted into separate wells in the gel. 10 à ¯Ã‚ Ã‚ ­l of the DNA ladder standard is pippeted into one well of each row on the gel. Running the gel The lid is place on the gel box, the electrode wires are connected to the power supply. The power supply is turned on to about 100 volts. To ensure the correct direction of the current, the movement of the blue loading dye is checked. The power supply is continued till the blue dye approaches the end of the gel. The wires are disconnected from the power supply. The lid is removed from the electrophoresis chamber. Using gloves, gel is carefully removed and observed in a transilluminator for the DNA bands. RESULTS: Isolation of DNA was done from blood ,fresh saliva, saliva stored at -20 °C, saliva stored at 37 °C for 24hrs ,buccal swab and betel quid by both the phenol-chloroform method and the chelex method. . Gel electrophoresis of the isolated genomic DNA was carried out on 0. 8% agarose gel. (figure 1) After restriction digestion electrophoresis gel is prepared to run and to identify the number of bands. DNA samplesobtained from blood were labelled as Aband subsequently as Bb,Cb, DbEbas shown in table 2. DNA obtained from fresh saliva were labelled as As,Bs, Cs, Ds,Es. DNA obtained from saliva stored at -20 degree were labeled as AfsBfsCfsDfs. Efs. DNA obtained from saliva stored at room temperature were labelled as Ads,BdsCdsDdsEds DNA obtained from bloodof 5 individuals was made to run in the well marked 1 to 5 in a uniform manner ie DNA obtained from the first individual named as Ab, was made to run in well No. 1 . DNA obtained from second individual named as Bb was made to run in well No. 2 DNA obtained from third individual named as Cb was made to run in well No . 3. DNA obtained from fourth individual named as Db was made to run in as well No- 4. DNA obtained from Fifth individual namedEbwas made to run in well No. 5. (table 2) But while running DNA obtained from saliva of different sources the order was changed randomly. For example DNA isolated from fresh saliva for the first individual (As) instead of being run in the first well ie well No -6 was made to run in the third well( well no 8) and DNA isolated from saliva stored at -20 degree for the first individual(Afs) instead of being run in the first well ie well No-11was made to run in the third well (well No. 13)and DNA isolated from saliva stored at room temperature for the first individual(Ads) instead of being run in the first well ie well No-16 was made to run in the fifth well (well No. 20). Likewise DNA isolated from different sources of saliva of different individuals made to run in different wells and the number of bands produced is identified . From the figure 1 it could be identified that the well number 1,8 ,13,20 corresponding to DNA isolated from the first individual from various sources named Ab AsAfs Ads identified by the yellow arrow has uniformly three bands. For the well number 2,7,14,19 corresponding to DNA isolated from the second individual from various sources named BbBsBfsBds identified by the blue arrow has uniformly 6 bands . various DNA isolated from the fifth individual from various sources namedEbEsEfsEds identified by the green arrow has uniformly 4 bands . From the above figure itcould be identified that the well number 1,10 corresponding to DNA isolated from different source for the first individual named Ab,AbS,identified by the yellow arrow has uniformly four bands. For the well number 2 and 6 corresponding to DNA isolated from second individual from blood and buccal swab named BbBbSl identified by the blue arrow has uniformly 6 bands . For the well number 3 and 7corresponding to DNA isolated from third individual from blood and buccal swab namedCbCbS identified by the red arrow has uniformly 5 bands . For the well number 4and 8 corresponding to DNA isolated from fourth individual from blood and buccal swab named DbDbs,identified by the aqua arrow has uniformly 7 bands. For the well number 5and 9 corresponding to DNA isolated from fifth individual from blood and buccal swab named EbEbsE identified by the green arrow has uniformly 8 bands . This shows that DNA obtained from an individual from blood and buccal swab produce uniform banding pattern. This shows that DNA obtained from an individual from various source produce uniform banding pattern . Identification of individual from traces of saliva which could be used for forensic application -Extraction of DNA from Buccal swab. Restriction digestion with Ecor-1 from extracted DNA obtained from above mentioned source has been done for identifying individuals. Blood was used as a control and compared with DNA bands from buccal swab. A total of 10 wells were created. DNA obtained from blood wer e labeled as Ab, Bb,Cb, Db,Ebas shown in tab 3. DNA obtained from Buccal swab were labeled as Abs,Bbs, Cbs, Dbs, Ebs. DNA obtained from blood from 5 individuals was made to run in the well marked 1 to 5 in a uniform manner. But while running DNA obtained from buccal swab the order was changed randomly. For example DNA isolated from buccal swab for the first individual (Abs) instead of being run in the first well ie well No -6 was made to run in the fifth well( well no 10). Likewise DNA isolated from buccal swab of different individuals was made to run in different wells and the number of bands produced is identified Different methods of DNA extraction is been followed in that, most widely used is phenol chloroform method . Many new methods of DNA extraction have been tried. The chelex method is one among then . To know the efficacy of the chelex method it was compared with that of phenol chloroform method. Of the two methods studied the chelex method proved to be more easy to handle and less time consuming in addition to yieds higher amount of DNA and is proved by quantification with U. V spectrometer as shown in fig. 2. DISCUSSION: Forensic odontology is a branch of forensics which analyses stains and organic liquids from the oral cavity or its contents, bite mark comparison, investigation of trauma and oral injuries such as personal injury cases, and dental malpractice. The fundamental requirement of a criminal investigation is that the victim and aggressor should be positively identified. Forensic dentistry aids in the forensic process by comparing the deceaseds dentition with that of previous dental records or by facilitating to shape the profile of an individual in terms of age at the time of death, sex and phylogeny to aid in identification. 11,12 Saliva has been a potential source of identification and is usually found in bite marks, cigarette butts, betel quid, postage stamps, envelopes and other objects. The first phase of the study intended to isolate DNA from saliva (under different conditions), by phenol-chloroform method and chelex method and compare the yield with that of blood . The second objective was to find out efficacy of these methods in extraction of DNA from traces of saliva ie from Buccal swab, and from Betel quid and which could be used for forensic application. 8 The presence of residues are considerably important as biological evidences, but forensic analysis of such evidences has been hindered by failures in extraction of human DNA. Consequently, it is indispensable in forensic science to establish a reliable method for extracting DNA from samples collected at the crime site. The most important objective was whether individuals can be identifed from samples of different source and to ascertain the applicability of the restriction digestion in forensics. 13,14 Blood was taken as control, saliva was divided into 3 parameters ie from fresh saliva, from saliva stored at -20 degree for24 hr from saliva stored in room temperature for 24 hr’s were obtained . Identification of individual has been done with restriction enzyme EcoRI. . The isolated DNA was digested using the restriction enzyme EcoRI(G|AATTC)The digested DNA was run on 1% agarose gel electrophoresis and the bands produced in each individuals DNA were scored and is proved that identification of individual can also be done by DNA fingerprinting or profiling. Agarose gel electrophoresis separates DNA fragments according to their size. The most important objective was whether individuals can be identified from samples of different source and to ascertain the applicability of the restriction digestion in forensics. 16 DNA fingerprinting is a technique that is used to represent like and unlike DNA that is present in different individuals. Nucleotide sequences which show significant variation from one individual to another are taken into consideration. 17The most important objective of the study was to ascertain whether individuals can be identified from samples of different source and to ascertain the applicability of the restriction digestion in forensics and the last objective was toCompare the DNA yield from manual and kit method. To prove that DNA could be extracted from traces of saliva , Buccal swab and Beetal quid was used . DNA could be extracted from buccal swab,beetal quid and quantification was done with U. V spectrometer. Comparison of DNA isolated from all the samples collected from all the individual using two different procedures has been done and comparison of yield of different sources showed the kit method to be more effective . Use of biological evidences like saliva, buccal swab and betel quid are compromised due to the quandary in extraction of human DNA. The present study had proved to establish a reliable method for extracting DNA from samples collected from different sources of saliva and from traces of salivary stains which was comparable to bloodin proving identification. Samples collected from different sources of saliva and from traces of salivary stains can also be assessed by DNA fingerprinting or profiling which is based on the fact that DNA is unique to every individual .

Comparing Relationships in E. M. Forster’s A Passage to India and Virg

Comparing Relationships in E. M. Forster’s A Passage to India and Virginia Woolf’s To the Lighthouse E. M. Forster’s A Passage to India and Virginia Woolf’s To the Lighthouse are concerned with the lack of intimacy in relationships. Forster’s novel is set in English-run India, the difference between race and culture being the center of disharmony. Woolf’s novel is set in a family’s summer house, the difference between genders being the center of disharmony. Despite this difference of scale, the disharmonies are much the same. Unity and intimacy are intertwined in both novels. Whereas the definitions of intimacy vary with each person, all of the characters strive for unity through their relations with others. The difference in ideas of intimacy are what prevent unity from being achieved. For the Indians, intimacy is a sharing of possessions and personal information that acknowledges equality. For the English, intimacy is similarity of background and allegiance. Thus, Heaslop tells his mother that he made a mistake by asking one of the Pleaders to sm oke with him because the Pleader then told all the litigants that he was in with the City Magistrate (Forster, 20). To the Pleader, this sharing of cigarettes and leisure time is an act of intimacy because it seems an acknowledgement of equality. To Heaslop, this is only a friendly act of social convention because equality is based on race and class, is something inherent, not given. The idea of intimacy as unity is a strain throughout A Passage to India. When Aziz thinks of his wife on the anniversary of her death, he wonders if he shall meet her in an afterlife, but does not have specific faith in an afterlife. He believes that â€Å"God’s unity was indubitable and indubitably ... ...ziz is frustrated that his attempt at conciliation is not successful. Unity requires intimacy because intimacy is an acknowledgement of equality. Only when one transcends limitations of gender and race, extends oneself beyond social codes that emphasize division can true unity be achieved. Both authors end their novels with an insinuation of a future that will be friendlier to intimacy and unity: Lily finally achieves unity in her painting and the final words of the land to Aziz and Fielding are â€Å"’No, not yet†¦No, not there.† (Forster, 282). Sometime, somewhere the English and the Indians will unite and man and woman will achieve gendered unity within the self. Works Cited Forster, E.M. A Passage to India. London: Everyman’s Library, 1991. Woolf, Virginia. To the Lighthouse. Introduction by D.M. Hoare, Ph.D. London: J.M. Dent and Sons Ltd., 1960.

Espana – creative writing

It was the first time I had seen a large wound, well the first I had remembered, and the first I would never forget. We were on holiday, the perfect holiday from my point of view. There were about twenty of us, varying in age, some from Birmingham, a couple from America but the majority from London. We were going on a road trip, ultimately to Spain but stopping on the way in Dijon, France. As incredible this holiday was starting to sound, the finest part to me was probably the fact that my parents were not here. I had come with my hyperactive 18 year old brother, my cousin who was a medical student, my two aunts, one of whom had brought her grandson along, my beloved nephew. I don't think I can remember a single moment where I wasn't laughing, smiling or simply enjoying the company of my family and the many friends we had made. The journey was astounding; it became one of those trips that could never be able to forget. Stopping in Dijon for a couple of days was amazing. It was the little things that made it unique, for instance my cousin, brother and I would take my nephew, Prem for walks in a stunning rose garden that was simply tranquil. He would want to be with us everywhere, reminding us of ourselves when we were younger and innocent, just he seemed to have five times as much energy as we did. Spain was an experience of wonder. From the people, the night life, foods, activities and that feeling I felt throughout the trip of sheer freedom. It seemed to be absolutely perfect, from the swimming pools to the beach nearby and the balconies that overlooked the entire city that just lit up at night. Being is such a magnificent place, it automatically made me get to know the people I had come with to a great extent. Having this great bond between us all, certainly made us experience Spain just that bit more. In fact, they had made an intense itinerary of things to do for the upcoming week; cram packed with everything you could possibly to in Spain. They had arranged places of great interest that sounded exceedingly pleasurable, but something else had caught my fascination and made be completely obsessed for the next week. It was the swimming pool, including the attached slides, diving facilities, an island with a connecting bridge and a general contemporary look that made it seem as if it was a pool from the next century. Addicted, is the only word that could describe me and my new passion for simply swimming for hours on end, feeding this new obsession of mine. It was here where I learnt how to do my first back flip dive, here where I found out that my swimming shorts could actually drop off if I was not careful, and here where I met my new friend Sarah. She had come to Spain every year for the past 6 years and being Flemish, communication was quite a task but it seemed worth it. We had become inseparable, giving me some of my most treasured memories, one of which was our incessant trips to the swings. I had a new blissful life full of ecstasy, one I would not want to ever give up. I still remember everyday day and moment of the idyllic time I had there. I can distinctively recall upon one day in particular. One I rather forget. It was the three days before I had to leave the resort and that morning, we got up and went straight into the swimming pool. Living life to the greatest, we were enjoying everything we possibly could. Doing our usual routine; Prem, Sarah and I were just taking pleasure in messing about in the pool. Sarah seemed to have gone swimming so much from her various holidays; she had learnt a few tricks. They were astonishing and she tried teaching them to me. Of course I wasn't the best at water gymnastics, but I tried hard and managed to learn some fragments! Soon enough I had got a decent trick of diving into the pool doing a half summersault. Ecstatic about my new accomplishment I forgot about Prem. He was at the back of my mind but to me he would just be playing around, kicking water while staying afloat from armbands. The next thing I remember was when I saw him, climb out of the pool, with some difficulty and strike a pose the way I did when I was just about to do my new diving trick. He shouted right across the pool to where I was and screamed, ‘Hey, Shradha look at me! ‘. That's when I froze. He definitely was not going to be able to make it and the edge of the pool was lined in a fashionable but extremely unpractical layer of sharp broken stones and sand merged together. In a matter of seconds I watched him jump, watched him scrape his shin deep as he twisted and watched him land in the water. At that point I swam as fast as I possibly could to him and jumped out of the pool, pulling him up as well. At this moment it hit me how no one we knew was here but had taken a trip to the beach, how I was left in charge of him and how I had been so irresponsible. As I examined his wound, I saw the deep gash in his leg between his knee and foot, and saw the blood trickling out. That was when a colossal wave of both guilt and worry hit me at once. Along with panic, it was a terrible combination. I was a mess of emotions that clouded my thinking and just made me confused, panicked and flustered. I was trying to comfort Prem and luckily Sarah ran into the hotel and called for help, which came quite fast. She had been the much more practical one, thinking ahead and trying to help the situation. I felt that I could not have been any more negligent and inattentive and that I was to blame for every tear that rushed down his face. The hotel staff came and got him in a relatively stable position but said that Prem should go get it cleaned up and bandaged properly. The hotel managed to contact my aunts and I didn't even know, as I was too wrapped up in other problems. In a matter of what felt like two minutes, my aunts came and saw Prem's leg. Being a doctor, my aunt inspected his leg and succeeded to rustle up a variety of things from the Hotel Staff and Manager. By now I had handed Prem over, and watched as he was being dealt with. I was simply amazed at his courage and bravery. He had stopped crying completely. Even when he was with me, I had not heard a complaint, moan or any sign of making things a hundred things worse than it was. I was learning so much from him, in such a short space of time. The whole incident seemed to have gone in a matter of minuets, but I had felt so many emotions flow through me. It was one of those days I would never forget, and from that day forward I understood so much more about my nephew. How he faced that day made me look at him from another light. For the next day, he complained about wanting to go swimming again. Also, when we got back to France, my cousin, brother and I would once again take my nephew, for walks in stunning rose and lilly gardens because he begged us to take him and never once did we refuse him.

The Jennifer Hudson Family Murder Case

On October 24, 2008, the bodies of Academy Award-winning actress Jennifer Hudsons mother and brother were found in the familys home on Chicagos South Side. Shot to death was Hudsons mother, Darnell Donerson, and her brother, Jason Hudson. Missing from the home was Julian King, the son of Jennifers sister Julia Hudson. Three days later the body of 7-year-old Julian, Hudsons nephew, was found in the back seat of an SUV parked on the West Side. He also had been shot. A .45-caliber gun found near the parked SUV was linked to all of the shooting deaths. The SUV was later confirmed to be that of Hudsons murdered brother, Justin King. A gun was also found in a vacant lot in the same neighborhood as the SUV, police said. The case drew national attention because of the fame of family member Jennifer Hudson, who won the best-supporting-actress Academy Award for her 2007 role in the film Dreamgirls. Hudson first gained fame after she was ousted on season three of the television talent show American Idol. Julias Estranged Husband Questioned William Balfour, the estranged husband of Julia Hudson, was taken into custody the day the first two bodies were found and held for 48 hours. He was then taken into custody by the Illinois Department of Corrections on a suspected parole violation. Balfour married Julia Hudson in 2006 but had been separated at the time of the shootings. He was thrown out of the Hudson home by Julias mother in the winter of 2007, according to reports. He denied any involvement with the Hudson case and denied statements that he had been seen with a gun, but remained in police custody. Balfour served almost seven years in prison after being convicted of attempted murder, vehicular hijacking and possession of a stolen vehicle. He was on parole at the time that the murder took place. Brother-in-Law Arrested Balfour was arrested at Stateville Correctional Center where he was being held on parole violation charges. Prosecutors believed that the shootings at the Hudson family home were the result of an argument Balfour had with Julia about another man. Investigators learned that Balfour tried to get a former girlfriend, Brittany Acoff-Howard, to provide him with a false alibi for the day that the murders occurred.   Im Going to Kill Your Family According to court records, Balfour threatened to kill members of Hudsons family on at least two dozen occasions before the three murders in October 2008. Assistant States Attorney James McKay said the threats began shortly after Balfour and his wife Julia Hudson broke up and he moved out of the family house. McKay said Balfour told Julia, If you ever leave me, Im going to kill you, but Im going to kill your family first. You will be the last to die. Jury Selection After answering questions about their knowledge of singer and actress Jennifer Hudson, 12 jurors and six alternates were chosen for the trial. Potential jurors in the trial were given questionnaires which asked if they were familiar with Hudsons career, if they regularly watched American Idol, and even if they were members of Weight Watchers, a weight-loss program for which Hudson is a celebrity spokesperson.   The jury was composed of 10 women and eight men and was racially diverse. While waiting for opening statements to begin a month later, Judge Charles Burns asked the jurors not to watch the television show American Idol, because Hudson was scheduled to make an appearance on an upcoming episode. The Trial During opening statements, Balfours defense attorney told jurors that police targeted him for the crime because they were under pressure to solve quickly what they knew would become a high-profile case, because of Jennifer Hudsons notoriety. Defense attorney Amy Thompson also told the jury that DNA found on the gun and fingerprints found in the SUV, in which Julians body was found three days later, did not match Balfour. Balfour pleaded not guilty to the charges and claimed he was nowhere near the house when the murders occurred. We Didnt Like How He Treated Her None of us wanted her to marry him [Balfour], Jennifer Hudson told the jury, We did not like how he treated her. Jennifer Hudsons sister Julia testified that Balfour was so jealous that he would even become angry when her son Julian kissed his mother. He would tell the 7-year-old, Get off my wife, she testified. Brittany Acoff Howard testified that William Balfour asked her to cover for him  for Oct. 24, 2008, the day Hudsons family members were killed.  Howard told jurors that Balfour helped buy her a prom dress and treated her like a little sister. He told me that if anybody asks you, Ive been out west all day, Acoff Howard said. In response to a specific prosecution witness, she said Balfour had asked her to lie for him. No DNA, But Gunshot Residue Illinois State Police evidence analyst Robert Berk told jurors that gunshot residue was found on the steering wheel of Balfours vehicle and the ceiling of the Suburban. His testimony followed that of another analyst, Pauline Gordon, who said no traces of Balfours DNA were found on the murder weapon, but that did not mean he never handled the gun. Some people shed skin cells faster, Gordon said. Gloves could have been worn. Guilty The jury deliberated 18 hours before finding Balfour guilty on three counts of murder and several other charges in connection with the October 24, 2008, deaths Darnell Donerson; Jason Hudson; and her 7-year-old nephew Julian King. After the verdict, jury members described the process they used during their almost 18 hours of deliberations. First, they voted on whether each witness was credible or not. Then they created a timeline of the crime to compare it with the alibi Balfours attorneys outlined during the trial. When the jury got around to taking its first vote, it was 9 to 3 in favor of conviction. Some of us tried our best to make him innocent, but the facts just werent there, juror Tracie Austin told reporters. Sentencing Before he was sentenced, Balfour was allowed to make a statement. In it, he offered condolences to the Hudson family but maintained his innocence. My deepest prayers go out to Julian King, Balfour said. I loved him. I still love him. Im innocent your honor. Under Illinois law, Balfour faced mandatory life without parole sentences for the multiple murders.  Illinois law does not allow death penalty sentences under any circumstances. You have the heart of an arctic night, Judge Burns told Balfour at his sentencing hearing. Your soul is as barren as dark space. Balfour was sentenced to life without parole. Grateful for Support Grammy and Academy Award-winning Hudson sobbed and leaned on her fiances shoulder as the jury verdict was read. She attended every day of the 11-day trial. In a statement, Jennifer and her sister Julia offered their gratitude: We have felt the love and support from people all over the world and were very grateful, the statement said. We want to extend a prayer from the Hudson family to the Balfour family. We have all suffered a terrible loss in this tragedy. They said they were praying that the Lord will forgive Mr. Balfour of these heinous acts and bring his heart into repentance someday. Balfour Continues to Deny Involvement In February 2016,  Balfour  talked was interviewed by  Chuck Goudie of  WLS-TV, ABC7s sister station in Chicago. This was his first publicized interview since his conviction. During the interview, Balfour stated that his conviction was due to a large conspiracy that included the police, witnesses, and lawyers and that he had nothing to do with the murders. When asked about why  7-year-old  Julian King was murdered, Balfours answer was chilling: Balfour: ...It could have been a wrong place at the wrong time, the person who come in there to kill somebody dont kill who they kill. If you are a witness and you can identify somebody, they can say I killed him because he could have identified me but thats not the case.Goudie: That 7-year-old boy could have identified you.Balfour: That what I said earlier, that he could identify me and thats why he got killed. Or he killed him because he could identify him. Now Julian was smart, he could remember faces. In response to the interview, the Chicago Police Department said: CPD stands firmly behind our investigation which was based exclusively on facts and evidence in this senseless murder. Balfour is currently serving his time in  Stateville Correctional Center  near  Joliet, Illinois.