DNA extraction of common garden broodstock. Eluted in 200 uL.
- NF3_17
- NF3_18
- NF3_19
- NF3_20
- NF4_1
- NF4_2
- NF4_3
- NF4_4
- NF4_5
- NF4_6
- NF4_7
- NF4_8
- NF4_9
- NF4_10
- NF4_11
- NF4_12
- NF4_13
- NF4_14
- NF4_15
- NF4_16
- NF4_17
- NF4_18
- NF4_19
- NF5_1
Did the ligation step for Plate 1 of the 2b-RAD broodstock libraries.
1st made fresh annealed adaptors.
|
Adaptor 1
|
96x
|
|
5ILL-NR
|
5
|
|
Anti-ILL
|
5
|
|
NFW
|
490
|
|
Adaptor 2
|
11x
|
|
3ILL -NR
|
5 uL
|
|
Anti-ILL
|
5 uL
|
|
NFW
|
490 uL
|
Removed 5 uL of digestion product from each well to give 10 uL.
|
1x
|
96x
|
|
|
2 uM Adaptor 1
|
5 uL
|
480
|
|
1 uM Adaptor 2
|
5 uL
|
480
|
|
T4 ligase
|
1 uL
|
96
|
|
T4 ligase buffer with 10 mM ATP
|
4 uL
|
384
|
|
NFW
|
24
|
2304
|
|
10 mM ATP
|
1
|
96
|
|
Total
|
40
|
3840
|
Let to digest at 16degC for 2 hours followed by 10 minutes at 62degC and then put in freezer.
In the evening did another set of extractions. These are repeats of the ones done on 10/27/15 that turned out poorly.
- SS4_1
- SS4_2 (messed up)
- SS4_3
- SS4_4
- SS4_5
- SS4_6
- SS4_7
- SS4_8
- SS4_9
- SS4_10
- SS4_11
- SS4_12
- SS4_13
- SS4_14
- SS4_15
- SS4_16
- SS4_17
- SS4_18
When vortexing SS4_2 the cap wasn’t on all the way so most of it splashed out. Need to re-extract.
Getting My Genes Wet 