Lab 6 - copies of a piece of DNA generating thousands to...

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DNA fingerprinting & PCR
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DNA fingerprinting Chemical structure of everyone’s DNA is same. Difference lies in base pairs Time consuming to look at millions of base pairs and sequence There are repeats in sequence Although 99.9% of human DNA sequences are the same in every person, enough of the DNA is different to distinguish one individual from another, unless they are monozygotic twins DNA profiling uses repetitive ("repeat") sequences that are highly variable, called variable number tandem repeats (VNTR), particularly short tandem repeats (STR)s.
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There are two principal families of VNTRs: microsatellites and minisatellites The former are repeats of sequences less than about 5 base pairs in length, while the latter involve longer blocks RFLP – Restriction digest followed by Southern blot to locate the VNTR loci Disadvantage – Laborious PCR Variations of VNTR allele in 6 individuals
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PCR (Polymerase Chain reaction) The polymerase chain reaction (PCR) is used to amplify a single or a few
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Unformatted text preview: copies of a piece of DNA generating thousands to millions of copies of a particular DNA sequence Initialization step : 9496 C which is held for 19 minutes. It is only required for DNA polymerases that require heat activation Denaturation step : 9498 C for 2030 seconds. It causes DNA melting of the DNA template by disrupting the hydrogen bonds between complementary bases, yielding single-stranded DNA molecules. Annealing step : 5065 C for 2040 seconds allowing annealing of the primers to the single-stranded DNA template. Elongation step : 7074 C Final elongation : This single step is occasionally performed at a temperature of 7074 C for 515 minutes after the last PCR cycle to ensure that any remaining single-stranded DNA is fully extended. Final hold : This step at 415 C for an indefinite time may be employed for short-term storage of the reaction....
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Lab 6 - copies of a piece of DNA generating thousands to...

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