Matt:LabNotes/2013-6-27

From ZhangLabWiki
Jump to navigation Jump to search

Continued from: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-26

Ethanol Precipitation after PAGE size selection[edit]

  • 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[edit]

  • Dilute 2ul into 20ul
  • Mix 1:1 with 2X buffer (20X dilution)
  • Load sample wells with 4,6,8,10,12 ul
  • Load Low Mass DNA ladders in 0.5, 1, and 2 ul
' 0.5 Ladder 1 Ladder 4 ul 6 ul 8 ul 10 ul 12 ul 2 Ladder
DNA volume 0.5 1 0.2 0.3 0.4 0.5 0.6 2
H2O 1.5 1 1.8 2.7 3.6 4.5 5.4 0
2X Buffer 2 2 2 3 4 5 6 2
Total 4 4 4 6 8 10 12 4

File:2013-06-27 0-gap-probes GelQuantification.jpg

  • 20X diluated sample concentration is 1.42 ng/ul
  • Starting sample concentration: 28.4 ng/ul -> total yield about 2.2ug/(193nt*325 Da/nt) = 35.1 pmole

Continued on: http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-6-28