Revision as of 21:18, 4 June 2013 by >Djacobse
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DpnII Digestion Test[edit]
Only running 1/4 of DNA I have for each sample to compare DpnII reaction yield using aadUTP DNA and normal DNA.
1. Set up
Sample
|
uL sample
|
uL Dpn II (50 U/uL)
|
uL 10x buffer
|
uL H20
|
total
|
alpha 4-A
|
35
|
2
|
5
|
8
|
50
|
alpha 6-A
|
35
|
2
|
5
|
8
|
50
|
beta 4-A
|
35
|
2
|
5
|
8
|
50
|
beta 6-A
|
35
|
2
|
5
|
8
|
50
|
2. Incubate in thermocycler at 37C for 2 hours; heat shock 20 minutes at 65 C
3. Column Purification, eluted with 30 uL EB
4. Stored at 4C; Gel running tomorrow
Nanodrop Results (from 6/4/2013)[edit]
Sample
|
ng/uL
|
ug in 30 uL
|
Yield (%)
|
alpha-4A
|
121.5
|
3.6
|
79.9
|
alpha-6A
|
136.7
|
4.1
|
78.1
|
beta-4A
|
126.4
|
3.8
|
73.7
|
beta-6A
|
129.7
|
3.9
|
75.7
|