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Cleanup each full pool via ultra purification
Size select for 250-350bp fragments size window via Pippin Prep
Sending Out for Sequencing to Admera
5% PhiX spike
Library 1
SAR_Plate1_Lib1
SAR_Plate3_Lib1
SAR_Plate5_Lib1
SAR_Plate7_Lib1
SAR_Plate9_Lib1
SAR_Plate11_Lib1
SAR_Under20_Lib1
Library 2
SAR_Plate2_Lib2
SAR_Plate4_Lib2
SAR_Plate6_Lib2
SAR_Plate8_Lib2
SAR_Plate10_Lib2
SAR_Plate12_Lib2
SAR_Over750_Lib2
Check In Samples Against List from Sam Patrick Johnson
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Dilute Super high tubes an additional 3x by adding 60 ul TE
High plate Plate 12 was diluted 4x by adding 60 ul TE; plate 3x by adding 40ul
Both plates and tubes were requantified
Normalize plates with TE on Nimbus
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Reagent | ul/rxn | rxns | ul needed (1.6x) | |
|---|---|---|---|---|
MseI oligo | 1 | 1600 | 1600 | 220 |
H2O | 0.112 | 1600 | 179.2 | 24.6 |
10x T4 Buffer | 0.1 | 1600 | 160 | 22 |
5M NaCl | 0.01 | 1600 | 16 | 2.2 |
1 mg/ml BSA | 0.05 | 1600 | 80 | 11 |
T4 DNA ligase | 0.1675 | 1600 | 268 | 37 |
Total | 1.4 | 1600 | 2240 | 220 |
Add 1 ul of EcoR1 Adaptors with 96-channel and Track which template plate gets which MID plate.
GTL Plate | EcoR1 MID plate | Pool Plate | Library | Norm to | Sequencer |
|---|---|---|---|---|---|
1Sauger1 | 1 | ||||
1Sauger3 | 2 | ||||
1Sauger5 | 3 | ||||
1Sauger7 | 4 | ||||
1Sauger9 | 5 | ||||
1Sauger11 | 6 | ||||
1Sauger2 | 1 | ||||
1Sauger4 | 2 | ||||
1Sauger6 | 3 | ||||
1Sauger8 | 4 | ||||
1Sauger10 | 5 | ||||
1Sauger12 | 6 | ||||
1SaugerUnderOver | 8 | ||||
Label reaction plates with MID plate used.
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Run Final Products on Tapestation for size check
1Sauger1
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1Sauger2
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