CTR:Notebook/Migrant/2016/08/23: Difference between revisions
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== | ==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 | |||
[[CTR:60px|60px]] | |||
1) 2uL 100bp low scale ladder, | |||
3) 2uL sheared TRBL | |||
→Sheared DNA to ~400bp. | |||
<br> | |||
* 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 dH<sub>2</sub>O and added 160 ul of mixture to each tube | |||
[[CTR:75px|75px]] | |||
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) | |||
**'''Assigned Adapter ''' | |||
*Gel of Final Library: | |||
[[CTR:60px|60px]] | |||
1) 2uL 100bp low scale ladder, | |||
3) 2uL TRBL | |||
* Picogreen: ng/μL | |||
*Run sample on BioAnalyzer DNA High Sensitivity Chip (2016-08-2) | |||
[[CTR:550px|550px]] | |||
*[ Bioanalyzer Results] | |||
*Sent 10nM to UC Davis for sequencing (Illumina HiSeq 250 PER (Rapid Mode)) (2016-08-2) | |||
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Revision as of 16:57, 24 August 2016
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Whole Genome Sequencing PCR-Free Library Prep for TRBL
1) 2uL 100bp low scale ladder, 3) 2uL sheared TRBL
→Sheared DNA to ~400bp.
1) 2uL 100bp low scale ladder, 3) 2uL TRBL
1) 2uL 100bp low scale ladder, 3) 2uL TRBL
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