Life Sci. 1B - Lab Quiz, (Human Origins 2)

Front 1

Describe binding of the PCR prdouct.

Back 1

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

Front 2

Describe washing of the PCR product.

Back 2

Ehtnaol-containing Buffer PE washes away only the primers and impurities, leaving the PCR product on the membrane

Front 3

Describe drying PCR product.

Back 3

Removes ethanol in the Buffer PE, which interferes with the sequences reaction.

Front 4

Describe eluting PCR product.

Back 4

Water releases the PCR rpdouct from the membrane

Front 5

What is the order to purify PCR?

Back 5

Binding, washing, drying, eluting

Front 6

What has to be done in order to sequence DNA?

Back 6

A sequences reaction will be set-up on a thermal-cycler

Front 7

Whta is the difference between sequence reaction and PCr?

Back 7

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

Front 8

Why do you set up 2 separate reactions for forward and reverse?

Back 8

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'

Front 9

Which primer wil be labeled the leading strand and which will be labeled the complement strand?

Back 9

F primer = leading strand
R primer = complement strand

Front 10

How many cycles are performed?

Back 10

30

Front 11

What are the corresponding colors for the DNA bases?

Back 11

Red = T
Blue = C
Black = G
Green = A

Front 12

Whta happens as the dye-labeled fragments migrate through the capillary array from the 5' end?

Back 12

The fluorescence is detected by laser excitation aand is recorded by a CCD camera.