Agarose Gel Prac 1 Flashcards
What was put into the wells
3 PCR tubes with fup rup and ip of the pblt101 insert
Then the 4 restriction tubes including the miniprep uncut
Which buffer filled the tank and also was used to dilute agarose
TAE
Why is tae important
Let’s dna run smooth in gel
What else was put in the agarose to see the dna
Nancy 520 - intercalating dna dye
What was used to produce wells
Comb
What was put into the samples and why
10 x loading buffer
Contains heavy glycerol to hold dna in tank
Has bromophenol blue dye so you know where samples are in wells
How do you work out the size (bp) of the u known bands
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
Why does super coiled circular dna run faster than linear or circular even if same bp size
It is more streamlined so moves faster
Why do circular dna run slower even if same bp amount
The conformation is harder to run on gel than linear or supercoiled
What can form between plasmids which makes them run slow on gel
Concatamers
Why did PCR 2 which had fup and ip not create a band
Both forward primers on same strand. Didn’t amplify 2 strands so didn’t show
