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...