Matt:LabNotes/2013-6-27

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Revision as of 16:46, 27 June 2013 by >Mzcai (Created page with "Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-26 ===Ethanol Precipitation after PAGE size selection=== *Spun 8 tubes in -80C at 10,000rp...")
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Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-26

Ethanol Precipitation after PAGE size selection

  • Spun 8 tubes in -80C at 10,000rpm at 4C for 30min
  • Discarded supernatant and added 650ul 75% EtOH to wash and spin at 14,000rpm at 4C for 5 min
  • Discarded supernatant and let dry in hood 10 min
  • Resuspended each of the 8 tubes with 10ul H2O (80ul total)

TBU Gel Quantification Plan

  • Dilute 2ul into 20ul
  • Mix 1:1 with 2X buffer (20X dilution)
  • Load sample wells with 2, 4, 6, 8 ul
  • Load Low Mass DNA ladders in 0.5, 1, and 2 ul
' 0.5 ladder 1 ladder 2 ul 4 ul 6 ul 8 ul 2 ladder
DNA volume 0.5 1 0.1 0.2 0.3 0.4 2
H2O 1.5 1 0.9 1.8 2.7 3.6 0
2X Buffer 2 2 1 2 3 4 2
Total 4 4 2 4 6 8 4
  • 20X diluated sample concentration is 1ng/ul
  • Starting sample concentration: 20ng/ul -> total yield about 1.2ug/(193nt*325 Da/nt) = 19.1 pmole