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12 Cards in this Set

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Describe binding of the PCR prdouct.
Buffer PB djusts the salt and pH optimum for DNA binding ~ silica membrane in the spin column absorbs only the PCR product under this condition
Describe washing of the PCR product.
Ehtnaol-containing Buffer PE washes away only the primers and impurities, leaving the PCR product on the membrane
Describe drying PCR product.
Removes ethanol in the Buffer PE, which interferes with the sequences reaction.
Describe eluting PCR product.
Water releases the PCR rpdouct from the membrane
What is the order to purify PCR?
Binding, washing, drying, eluting
What has to be done in order to sequence DNA?
A sequences reaction will be set-up on a thermal-cycler
Whta is the difference between sequence reaction and PCr?
1. You use PCR produt as a NDA template
2. dNTPs are fluorescently labeled
3. includes only one of the primer pair at a time
Why do you set up 2 separate reactions for forward and reverse?
You want to amplify so that only one of the strands is fluorescently labeled inone reaction. Both the forward and reverse primers will albel from 5' to 3'
Which primer wil be labeled the leading strand and which will be labeled the complement strand?
F primer = leading strand
R primer = complement strand
How many cycles are performed?
What are the corresponding colors for the DNA bases?
Red = T
Blue = C
Black = G
Green = A
Whta happens as the dye-labeled fragments migrate through the capillary array from the 5' end?
The fluorescence is detected by laser excitation aand is recorded by a CCD camera.