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Lambda Exonuclease Digestion[edit]
Reaction Table
Reagent
|
uL
|
Amplicon
|
70
|
Lambda Exonuclease Buffer (10X)
|
10
|
Lambda Exonuclease
|
5
|
nf H2O
|
15
|
Total
|
100
|
- Set up reaction according to table
- Incubate at 37C for 1 hour
- Purify using 2 ssDNA columns, elute with 20 uL nfH2O
Results[edit]
Sample
|
ng/uL
|
ug in 20 uL
|
ug Total
|
PPRL-XO-A
|
10.5
|
0.2
|
|
PPRL-XO-B
|
20.8
|
0.4
|
0.6
|
So there is very little left. I'm going to do a bigger amplification tomorrow, and I'll pool today's samples with tomorrow's before continuing with USER/DpnII.