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

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...