Revision as of 02:35, 12 May 2013 by >Djacobse
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Ethanol Precipitation
7. Centrifuged 10000 rpm for 30 minutes at 4C
8. Removed supernatant added 750 uL 75% EtOH
9. Centrifuged 10000 rpm for 12 minutes at 4C
10. Removed supernatant, dried in hood
11. Resuspended with 50 uL
12. Column Purification, eluted with 50 uL
13. measured concentration in nanodrop
Nanodrop Results
Sample
|
V4A
|
V4B
|
V4C
|
V4D
|
V6A
|
V6B
|
V6C
|
V6D
|
ng/uL
|
127.5
|
156.3
|
130.2
|
130.6
|
142.7
|
138.9
|
146.4
|
133.4
|
ng
|
5100
|
6252
|
5208
|
5224
|
5708
|
5556
|
5856
|
5336
|
ug in 4 sets
|
|
|
|
21.8
|
|
|
|
22.5
|
DpnII Digestion
Enzyme Info
1. Prepared samples according to table
*DpnII-Lot 0091201; Buffer 0031105
Sample
|
V4A
|
V4B
|
V4C
|
V4D
|
V6A
|
V6B
|
V6C
|
V6D
|
ng/uL
|
127.5
|
156.3
|
130.2
|
130.6
|
142.7
|
138.9
|
146.4
|
133.4
|
ng
|
5100
|
6252
|
5208
|
5224
|
5708
|
5556
|
5856
|
5336
|
uL
|
35
|
35
|
35
|
35
|
35
|
35
|
35
|
35
|
uL 10x buffer
|
10
|
10
|
10
|
10
|
10
|
10
|
10
|
10
|
uL DpnII (50U/uL)
|
2
|
2
|
2
|
2
|
2
|
2
|
2
|
2
|
uL nf H20
|
53
|
53
|
53
|
53
|
53
|
53
|
53
|
53
|
Total
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
100
|
2. Incubated 16 hours at 37C, followed by 20 minutes at 65C