Blue:RNA-Seq Experiments:02272014: Difference between revisions
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>B1lake (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 (→Tests) |
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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