Blue:RNA-Seq Experiments:02272014: Difference between revisions

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(Created page with "== Human Nuclei C1 Experiment: 02-27-2014 == == Purpose == *To test conditions for preventing Nuclei clumping during C1 loading == Tests == *Sort 50K nuclei per tube in B...")
 
>B1lake
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== Tests ==
== Tests ==


*Sort 50K nuclei per tube in BSA buffer
*Sort 50K nuclei per tube in 1% BSA buffer
*Centrifuge 910g at 4C for 10 min
*Centrifuge 910g at 4C for 10 min
*Resuspend in BSA sort buffer containing:
*Resuspend in 1% BSA sort buffer containing:
:*No supplements
:*No supplements
:*1mM MgCl2
:*1mM MgCl2

Revision as of 19:25, 27 February 2014

Human Nuclei C1 Experiment: 02-27-2014

Purpose

  • To test conditions for preventing Nuclei clumping during C1 loading


Tests

  • Sort 50K nuclei per tube in 1% BSA buffer
  • Centrifuge 910g at 4C for 10 min
  • Resuspend in 1% BSA sort buffer containing:
  • No supplements
  • 1mM MgCl2
  • 25% Glycerol
  • 10% PEG6000
  • 1mM MgCl2/25% Glycerol
  • 1mM MgCl2/10% DMSO
  • Count using Biorad T20
  • Add 7ul (containing 800 nuclei) to 3ul C1 Suspension RGT
  • Incubate 10 min at RT
  • Centrifuge @910G for 10 min
  • Gently resuspend and add into BioRad slide for counting/sizing