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4.2 - B No 5’3’ exonuclease C Little or no 3’ 5’...

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I. ddNTP sequencing A. yield is about 750 bp per run B. high throughput is acheievd by increasing capacity C. very expensive D. achron x prize, 10 million to first team that can sequence 100 genomes within 10 days at a cost of no more than 10,000 dollars II. The sequencing template must be single stranded A. double strand plasmind SNA is denatured with heat or alkali B. the library is created in a vector that can be converted to a single stranded form ex M13 bacteriophage of a plasmid containing the M13 ori III. DNA polymerase for sequencing A. High porcessivity B. No 5’3’ exonuclease
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Unformatted text preview: B. No 5’3’ exonuclease C. Little or no 3’ 5’ exo nuclease – would tend to remove ddNTP which terminates the fragments you want to detect D. Modified T7 DNA polymerase mutation have eliminated exonuclease activit, called sequenase IV. Thermostable DNA polymerase A. no PCR because you are using only 1 primer B. linear increase in amount of DNA, not exponential (like in PCR) C. advantages 1. dbl ss DNA is used bc template is melted in each heating cycle 2. very little template needed 3. chain terminated strands increase linearly...
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