CTR:Notebook/Migrant/2015/09/29

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Whole Genome Sequencing PCR-Free Library Prep for YWAR & WIFL Used TruSeq Library Prep Kit Diluted samples 77585 (YWAR) & 1590-97213 (WIFL) to 1ug of DNA in 100 uL total of Buffer AE Sonication (2015-09-29) BioRuptor NGS: 4 cycles of 30 seconds on, 90 seconds off, High Power 1) 2uL sheared YWAR 2) 2uL 100bp low scale ladder, 3) 2uL sheared WIFL →Sheared DNA to ~400bp. Dried down samples to get concentrated volume of 50 uL using SpeedVac (2015-09-29) Clean-up fragmented DNA (2015-09-30) End Repair and Size Selection- edited to size select for 400bp (2015-09-30) Made a diluted bead mixture with the ratio of 105ul beads: 79ul dH2O and added 160 ul of mixture to each tube 1) 2uL YWAR 2) 2uL 100bp low scale ladder, 3) 2uL WIFL Adenylate 3' Ends (2015-09-30) Ran on JOHN thermocycler under program: SDCRKAD, Block B Ligate Adapters (2015-09-30) Assigned Adapter 2 to YWAR and Adapter 4 to WIFL Gel of Final Library: 1) 2uL YWAR 2) 2uL 100bp low scale ladder, 3) 2uL WIFL Picogreen: YWAR: 7.42 ng/μL WIFL: 6.72 ng/μL Run 1μL of sample + 2μL of dH2O on BioAnalyzer DNA High Sensitivity Chip (2015-09-30) YWAR WIFL Bioanalyzer Results Sent 10nM to Berkeley for sequencing (Illumina HiSeq 250 PER (Rapid Mode)) (2015-10-01)