Rui:SingelCell: Difference between revisions

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==Protocols==
==Protocols==
===Single cell RNA isolation===
===Single cell RNA isolation===
*Single cell capture
**Maintain the integrity of living single cells before lysis (dissociation - transfer - wash - transfer)
**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.
**PBS alone will cause tight attachment of cells to dish/micropipette, leading to rupture/loss of cells.
 
===PCR-based protocol===
===PCR-based protocol===
===IVT-based protocol===
===IVT-based protocol===

Revision as of 20:57, 9 January 2011

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

  • 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.

PCR-based protocol

IVT-based protocol

Circulation-based protocol

2011

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