CTR:Notebook/Migrant/2016/08/23: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 37: Line 37:


*Ligate Adapters (2016-08-25)
*Ligate Adapters (2016-08-25)
**'''Assigned Adapter '''
**'''Assigned Adapter: 9'''


*Gel of Final Library:
*Gel of Final Library:

Revision as of 12:15, 25 August 2016

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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)
    • Assigned Adapter: 9
  • Gel of Final Library:

60px

1) 2uL 100bp low scale ladder,

3) 2uL TRBL

  • Picogreen: ng/μL
  • Run sample on BioAnalyzer DNA High Sensitivity Chip (2016-08-2)

550px

  • [ Bioanalyzer Results]
  • Sent 10nM to UC Davis for sequencing (Illumina HiSeq 250 PER (Rapid Mode)) (2016-08-2)
Retrieved from "https://openwetware.org/mediawiki/index.php?title=CTR:Notebook/Migrant/2016/08/23&oldid=946658"