Whole Genome Sequencing PCR-Free Library Prep for TRBL
- Used TruSeq Library Prep Kit
- Diluted sample 02N9685 to 1ug of DNA in 100 uL total of Buffer AE
- Sonication (2016-08-23)
- BioRuptor NGS: 4 cycles of 30 seconds on, 90 seconds off, High Power
1) 2uL 100bp low scale ladder,
3) 2uL sheared TRBL
→Sheared DNA to ~400bp.
- Dried down samples to get concentrated volume of 50 uL using SpeedVac (2016-08-23)
- Clean-up fragmented DNA (2016-08-24)
- End Repair and Size Selection- edited to size select for 400bp (2016-08-24)
- Made a diluted bead mixture with the ratio of 109ul beads: 75ul dH2O and added 160 ul of mixture to each tube
1) 2uL 100bp low scale ladder,
3) 2uL TRBL
- Adenylate 3' Ends (2016-08-25)
- Ran on JOHN thermocycler under program: SDCRKAD, Block B
- Ligate Adapters (2016-08-25)
- Run sample on BioAnalyzer DNA High Sensitivity Chip (2016-09-12)
- Sent 10nM to UC Davis for sequencing (Illumina HiSeq 250 PER (Rapid Mode)) (2016-09-13)
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