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MIT 7 014 - Recombinant DNA—Forensic

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Recitation Section 20Recombinant DNA—ForensicsSolve a mysteryRecitation Section 20 May 3-4, 2006 Recombinant DNA—Forensics Solve a mystery BARNEY AND THE BEAVER ARE GONE!! Help us solve the mystery so Professor Walker can continue to bring his trusted visual aids to 7.014. FACTS: • Barney and the beaver were seen by all in lecture on Wednesday, February 8. • The last time Barney and the beaver were seen was at a staff meeting in Professor Walker's office immediately after that lecture. • Jerry, Alicia, Lily, Julia, and Professors Klopfer and Walker were all present at the meeting. • One of these staff members (not Professor Walker) took the props. The techniques of dideoxy DNA sequencing and the polymerase chain reaction (PCR) were used to solve the crime. Cells left on the ransom note were isolated and the region containing gene K amplified by PCR. The DNA sequence of the PCR products was then determined. Shown in the box below are the results from a sequencing gel of a region of gene K. ddG ddA ddT ddC-+ 1. Write the DNA sequence for this region of gene K as shown in the sequencing gel above. Indicate the 5' and 3' ends of the DNA sequence. 2. One of the suspects, Jerry, has the following ds DNA sequence for this same region in both copies of gene K: 5' CTGAAGTACGT 3' 3' GACTTCATGCA 5' Is Jerry still considered a suspect in the case? Justify your answer. 1Analysis of a second site, the gene L locus (represented by the shaded box below), is useful for forensics. The only difference between the two alleles at this locus is that the l allele lacks an internal EcoRI restriction site present in the L allele. EcoRIallele:L allele:l PCR analysis of cells taken from the note indicated that it was handled only by a Ll individual. Based on this, the gene L locus of the suspects was amplified by PCR, the PCR products were digested with EcoRI, and the resulting fragments were separated by agarose gel electrophoresis as shown below: Larger frag m ent sSm aller frag m ent s–+genotype:Marissa Ad a m FangAlex Jessica Prof. Klopfer Julia Alicia Jerry Lily 3. In the spaces provided above, indicate the genotypes at the gene L locus for these suspects. 4. Based on the PCR analysis of the gene L locus, which individual(s) can be excluded as suspects in the crime? Justify your answer. At chromosomal position Z, it is common to find one or more copies of a 100 bp insert. Fortunately, most of the suspects have a different number of inserts at this position. We even tested Professor Walker to be sure: Prof. Klopfer has no inserts. Jerry has one insert. Lily has two inserts. Alicia has two inserts. Julia has three inserts. Professor Walker has four inserts. A partial DNA sequence of position Z is given below. The length of position Z without an insert is 2000 bp. 5'-GTGCA.... 3'-CACGT........CGACG-3'....GCTGC-5'[]100 bp insertn 25. To determine who took Barney and the beaver, design a PCR-based strategy to analyze position Z, including the sequences and orientation (5' and 3' ends) of two 5-base primers necessary to PCR amplify this region. 6. The following pattern was seen on the gel used to seperate the PCR products from the above experiment. 25002400230022002100200012 3 4 56 Molecular marker Sample from the ransom note i) Which lane corresponds to which individual? ii) Who committed this heinous crime?


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MIT 7 014 - Recombinant DNA—Forensic

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