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

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List of needed supplies to carry out the listed protocols:

Clean RNA technique

  • DEPC or DMPC (DMPC is less carcinogenic) for making RNAse free solutions/containers. --> Bought from Sigma 6.23
  • Peroxide --> bottles of unknown age, but chemical is still reactive. might as well use, since it's only used as a cleaning agent.
  • Deionized glyoxal --> Bought from Sigma 6.23

Sean Moore's RNA Extraction Protocol

  • rna- strain of Top10 --> If we can get this great, if not, no worries.
  • BPER lysis soln --> Sean says this is optional but still uses is as his solvent. We have this.
  • MgOAc --> on RTS shelf
  • Lysozyme --> can't find, let's buy more from Pierce I believe (BC)
  • DNAaseI --> NEB stuff in shared enzymes box. Buffer conditions differ from Sean's lysis soln, but should be OK.
  • Linear polyacrylamide (can't contain bis) --> bought from Sigma 6.23
  • Ambion's phenol/choloroform mix (pH 4.5) --> bought from Ambio 6.23

Northern blotting (Endy) Protocol

  • SybrGold --> have some aliquots in Barry/Jason's -20
  • BPTE buffer:
    • PIPES (RTS shelf)
    • Bis-Tris (RTS shelf)
    • EDTA (DE shelf)
  • RNA loading buffer:
    • 95% deionized formamide
    • 0.025% Bromophenol blue (solid on RTS shelf)
    • 0.025% xylene cyanol FF (solid on RTS shelf)
  • TurboBlotter apparatus --> asked Heather where this is.... waiting for reply
  • Nytran SuPerCHarge or Immobilon NY+ nylon membranes? --> might ask Sean what works better, or if it even matters...
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