User:Barry Canton/Notebook/T7 RNAP transcription of rRNA/2008/06/20: Difference between revisions

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*Protocol for Northerns
*Protocol for Northerns
*Equipment needed to image gel
*Equipment needed to image gel
*Relative vs "quantitative"


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Revision as of 12:10, 20 June 2008

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Things to do

  • Purchase reagents for RNA extraction via Sean's protocol
  • Purchase reagents for Northern blotting according to Brian's protocol
  • Grow cells
  • Lyse (denaturing lysis is fine)
  • How do we store lysates
  • Design and order probes
  • Design and order a standard to compare hybridization efficiency.
  • Do test Northerns to check primer.
  • Decide if we should try to get quantitative measures of rRNA levels or simply pursue measurements relative to wild-type rRNA levels.

Timeline

  • Order reagents today
  • Do extractions Tuesday/Wednesday
  • Do trial Northern Thursday
  • Try polysome stuff the following week

Questions for Brian

  • Any problems with RNAse when doing the native lysis?
  • Protocol for Northerns
  • Equipment needed to image gel
  • Relative vs "quantitative"


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