User:Barry Canton/Notebook/T7 RNAP transcription of rRNA

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Introduction
I have been transcribing rRNA with T7 RNA polymerase in E. coli. I would like to estimate how much immature rRNA transcript I am producing relative to that being made on the chromosome. I would also like to estimate how efficiently the T7-transcribed rRNA is being incorporated into active, functional ribosomes.

Milestones

 * 1) Specific detection of wild-type and mutant 16s rRNA
 * 2) *Design 3 sets of primers (use Bryan's design first). Order Monday
 * 3) *Do extraction on Wednesday?
 * 4) *Do blot on Wednesday/Thursday.
 * 5) *Pick best primer pairs
 * 6) Relative quantification of wild-type and mutant 16s rRNA
 * 7) Evaluation of efficiency of processing of mutant rRNA relative to wild-type rRNA.
 * 8) Write & Submit paper.

Protocols

 * Sauer:RNA Purification from E. coli
 * Sean's RNA Purification protocol - short
 * Northern Blotting
 * Polysome gradient running
 * List of Supplies

Related protocols

 * Native and denaturing lysis protocols collected from OWW.
 * P1 transduction to make the rna knockout


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