Rui:SingelCell: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>RuiLiu
>RuiLiu
m (Blanked the page)
 
(140 intermediate revisions by 3 users not shown)
Line 1: Line 1:
==Background==
*Goal: the smallest functional unit; the exact quantity; full-length RNAs; sense vs anti-sense; all classes; at single-base resolution


*Step 1: single cell; exact quantity; the 3' end; mRNA; RNAseq
==Protocols==
===Single cell RNA isolation===
1. Single cell capture
Maintain the integrity of living single cells before lysis (dissociation - transfer - wash - transfer)
PBS alone will cause tight attachment of cells to dish/micropipette, leading to rupture/loss of cells.
2. Cell lysis
Mild lysis buffer to ensure efficient lysis (thoroughly and completely lysed) w/o severely interfering with RT efficiency
Negative control to rule out the contamination from the start
===PCR-based protocol===
===IVT-based protocol===
===Circulation-based protocol===
==2011==
<calendar>
name=Rui:LabNotes/SingleCell
format=%name/%year-%month-%day
date=2011/10/1
view=oneyear
</calendar>

Latest revision as of 00:51, 7 December 2011