AlanFung:LabNotes/Sequencing/2009-11-11: Difference between revisions

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(New page: ==Rescue CVF-N==)
 
>Alan6017518
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==Rescue CVF-N==
==Rescue CVF-N==
*20ul of CVF-N library, with buffer added instead of iProof master mix
==Ethanol Precipitation==
*Mix 50ul 100% ethanol, 2ul 3M NaoAC, and 1ul glycoblue
*Store at -80C for 20min
*Centrifuge 10,000 for 20min at 4C
*Discard supernatant
*Add in 750ul 75%ethanol
*Spin 10,000 rpm for 5 min at 4C
*Discard supernatant
*let dry in hood for 10 min
*Add in 21ul ddh2o
Repeat PCR of sequencing library with the following sets

Revision as of 20:36, 11 November 2009

Rescue CVF-N

  • 20ul of CVF-N library, with buffer added instead of iProof master mix

Ethanol Precipitation

  • Mix 50ul 100% ethanol, 2ul 3M NaoAC, and 1ul glycoblue
  • Store at -80C for 20min
  • Centrifuge 10,000 for 20min at 4C
  • Discard supernatant
  • Add in 750ul 75%ethanol
  • Spin 10,000 rpm for 5 min at 4C
  • Discard supernatant
  • let dry in hood for 10 min
  • Add in 21ul ddh2o
Repeat PCR of sequencing library with the following sets