Revision as of 23:10, 9 August 2013 by >Djacobse
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3. Column purification, elute with 30 uL EB
4. Measure in Nanodrop
Nanodrop Results
Sample
|
V4S2AB
|
V4S2CD
|
V6S2A
|
V6S2B
|
V6S2C
|
V6S2D
|
ng/uL
|
109.3
|
122.9
|
115.4
|
117
|
121
|
111.1
|
ug in 30 uL
|
3.3
|
3.7
|
3.5
|
3.5
|
3.6
|
3.3
|
Total ug
|
|
7.0
|
|
|
|
13.9
|
Gel Image
File:2013-8-9-DpnIIDigest.jpg
Lambda Exonuclease Digestion
1. Reaction Table
Sample
|
V4S1C
|
V4S1D
|
V6S1A
|
V6S1B
|
V6S1C
|
V6S1D
|
ng/uL
|
109.3
|
122.9
|
115.4
|
117
|
121
|
111.1
|
ug in 25 uL
|
2.7
|
3.1
|
2.9
|
2.9
|
3.0
|
2.8
|
uL added
|
25
|
25
|
25
|
25
|
25
|
25
|
uL 10x buffer
|
5
|
5
|
5
|
5
|
5
|
5
|
uL Lambda Exonuclease (5U/uL)
|
3
|
3
|
3
|
3
|
3
|
3
|
uL nfH2O
|
17
|
17
|
17
|
17
|
17
|
17
|
total
|
50
|
50
|
50
|
50
|
50
|
50
|
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
3. Pool samples
4. ssDNA column purification, elute with 20 uL
5. Measure Concentration in Nanodrop
Nanodrop Results
|
ng/uL
|
ug in 20 uL
|
V4S2
|
94.7
|
1.9
|
V6S2
|
195.2
|
3.9
|