3A.Below are a few (of the many) possible errors that could adversely affect PCR or analysis of PCR products.  Explain why these errors are bad and predict if the error would lead to no product or just a low yield.  i)  Primer and DNA volumes reversed (2.5 µl primer and 22.5 µl cheek DNA) ii)  Student is homozygous for a mutation that deletes the four bases that would normally anneal to the last four bases at the 3´end of the primer  iii)  The PCR tube was not capped tightly (prior to the tube being placed in the thermal cycler) iv) While preparing a sample of the PTC PCR reaction to run on a gel, you intend to pipet up 8 µl of FD1 loading dye, but inadvertently depress the pipet plunger to the second stop instead of the first stop.  How would this pipetting error affect your conclusions from the Flash Gel?  Note: the error was made while preparing the sample with dye not while loading the gel.

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
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3A.Below are a few (of the many) possible errors that could adversely affect PCR or analysis of PCR products.  Explain why these errors are bad and predict if the error would lead to no product or just a low yield. 

i)  Primer and DNA volumes reversed (2.5 µl primer and 22.5 µl cheek DNA)

ii)  Student is homozygous for a mutation that deletes the four bases that would normally anneal to the last four bases at the 3´end of the primer 

iii)  The PCR tube was not capped tightly (prior to the tube being placed in the thermal cycler)

iv) While preparing a sample of the PTC PCR reaction to run on a gel, you intend to pipet up 8 µl of FD1 loading dye, but inadvertently depress the pipet plunger to the second stop instead of the first stop.  How would this pipetting error affect your conclusions from the Flash Gel?  Note: the error was made while preparing the sample with dye not while loading the gel.

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