LS4_11_MolGen2

LS4_11_MolGen2 - LS4 Final Exam ! announcements Thursday,...

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LS4 Final Exam announcements Thursday, Sept 10, 8:00-11:00 a.m. Sections 2B, 2C, 2D in MS 4000A TA: Aditi, Deepthi (2C) Sections 2A, 2E, 2F in CS 76 TA: Mohsin, Deepthi (2E)
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Must have photo ID. NO GRAPHING CALCULATORS!!! NO drinks, food, hats. No cell phones; all personal items to the front of the room. Emergency bathroom visits will be escorted. No other questions answered while only 1 proctor in the room. LS4 Final Exam announcements
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DNA Fingerprinting With the exception of identical twins or other clones, each organism has a unique DNA sequence. The two classes of repeat elements can be used to generate a DNA pattern or “fingerprint” that can identify specific individuals. Simultaneous comparison of the genotypic patterns of molecular markers at many different loci can determine the relationship between different individuals. Useful in criminal cases and to determine parentage.
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Any molecular marker can be used for DNA fingerprinting. RFLP , minisatellites (VTNR), microsatellites… - VNTR (variable number of tandem repeats) =minisatellite GCGGACCTATTAAGA GACCTATTAAGAGACCTATTAAGA… - STR (short tandem repeat)= microsatellite ATGCGGT CATCATCATCATCAT TTGACCT (CAT)n DNA Fingerprinting
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•Variation in size is due to a variable number of repeats , not to a change in the sequence of a restriction site. •Average size of repeats is 2-15kb. •Found very frequently in genomes. Minisatellites (VNTRs)
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1 1’ 200 bp 180 bp Microsatellites: polymorphic tandem repeats (2-8 bp long) CA, AT, CGA, etc. analyzed by PCR (CA)20 (CA)10
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Paternity Testing
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Detection of SNPs R estriction f ragment l ength p olymorphisms (RFLPs) - Restriction Digestion/Southern blot • Allele specifc oligonucleotides (ASOs) - PCR ±ollowed by blotting • DNA sequencing
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• Allele specific oligonucleotides (ASOs)…PCR followed by blotting: Detection of SNPs: ASOs
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• Allele specifc oligonucleotides (ASOs)… What are the genotypes of the following individuals? Detection of SNPs: ASOs A: Normal ß-globin allele S: Sickle ß-globin allele 1. Blot DNA from each individual twice (2 rows) 2. Probe each row with different ASOs. Probe for normal allele (A) Probe for Sickle allele (S)
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POSITIONAL CLONING Process of determining a gene from a mutant phenotype.
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This note was uploaded on 09/09/2009 for the course LS LS 4 taught by Professor Ribaya during the Spring '07 term at UCLA.

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LS4_11_MolGen2 - LS4 Final Exam ! announcements Thursday,...

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