Agarose Gel Prac 1

Question 1

What was put into the wells

Answer 1

3 PCR tubes with fup rup and ip of the pblt101 insert

Then the 4 restriction tubes including the miniprep uncut

Question 2

Which buffer filled the tank and also was used to dilute agarose

Answer 2

TAE

Question 3

Why is tae important

Answer 3

Let’s dna run smooth in gel

Question 4

What else was put in the agarose to see the dna

Answer 4

Nancy 520 - intercalating dna dye

Question 5

What was used to produce wells

Answer 5

Comb

Question 6

What was put into the samples and why

Answer 6

10 x loading buffer

Contains heavy glycerol to hold dna in tank
Has bromophenol blue dye so you know where samples are in wells

Question 7

How do you work out the size (bp) of the u known bands

Answer 7

Do distance in mm of each of the molecular ladder bands over x which is log 10 of the bp length

To find the bp length intercept the y by measuring mm of the band to find x

Question 8

Why does super coiled circular dna run faster than linear or circular even if same bp size

Answer 8

It is more streamlined so moves faster

Question 9

Why do circular dna run slower even if same bp amount

Answer 9

The conformation is harder to run on gel than linear or supercoiled

Question 10

What can form between plasmids which makes them run slow on gel

Answer 10

Concatamers

Question 11

Why did PCR 2 which had fup and ip not create a band

Answer 11

Both forward primers on same strand. Didn’t amplify 2 strands so didn’t show