User:Barry Canton/Notebook/T7 RNAP transcription of rRNA/List of Supplies: Difference between revisions
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==Sean Moore's RNA Extraction Protocol== | ==Sean Moore's RNA Extraction Protocol== | ||
*rna- strain of Top10 | *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 | *BPER lysis soln --> Sean says this is optional but still uses is as his solvent. We have this. | ||
*MgOAc --> on RTS shelf | *MgOAc --> on RTS shelf | ||
*Lysozyme --> can't find, let's buy more from Pierce I believe | *Lysozyme --> can't find, let's buy more from Pierce I believe |
Revision as of 10:12, 23 June 2008
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. --> none in Endylab
- 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 --> have an old bottle, but quality is questionable.
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
- DNAaseI --> NEB stuff in shared enzymes box. Buffer conditions differ from Sean's lysis soln, but should be OK.
- Linear polyacrylamide --> Samantha thinks this might be what we have, but we should ask Sean to make sure.
- Ambion's phenol/choloroform mix --> need to ask Sean which one, since Ambion sells several types
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...