Sauer:Protocols: Difference between revisions
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'''PLEASE NOTE:''' Blindly following a protocol is not a good substitute for actually talking to someone who has done the experiment. When posting protocols, please also include the name(s) of people in the lab who will be good to talk to about the protocol. | '''PLEASE NOTE:''' Blindly following a protocol is not a good substitute for actually talking to someone who has done the experiment. When posting protocols, please also include the name(s) of people in the lab who will be good to talk to about the protocol. | ||
==Protein Expression/Purification== | |||
[[Sauer:ClpX purification]] | [[Sauer:ClpX purification]] | ||
[[Sauer:ClpP purification]] NEEDS TO BE FILLED IN, WORK ON ME! | [[Sauer:ClpP purification]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[Sauer:Expression/purification of 35S-Met proteins]] | |||
==''In vitro'' enzymatic assays== | |||
[[Sauer:In vitro degradation of GFP-ssrA by ClpXP]] | [[Sauer:In vitro degradation of GFP-ssrA by ClpXP]] | ||
[[Sauer:In vitro degradation of radiolabeled proteins by ClpXP]] NEEDS TO BE FILLED IN, WORK ON ME! | [[Sauer:In vitro degradation of radiolabeled proteins by ClpXP]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[Sauer:ATPase assays]] NEEDS TO BE FILLED IN, WORK ON ME! | [[Sauer:ATPase assays]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
==Analysis of proteins or RNA ''in vivo''== | |||
[[Sauer:Western blot]] NEEDS TO BE FILLED IN, WORK ON ME! | [[Sauer:Western blot]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
| Line 22: | Line 27: | ||
[[Sauer:Quantitative Western blot]] NEEDS TO BE FILLED IN, WORK ON ME! | [[Sauer:Quantitative Western blot]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[Sauer: | [[Sauer:Northern blot]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[Sauer: | [[Sauer:In vivo tmRNA tagging assays]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[Sauer: | [[beta-Galactosidase Assay (A better Miller)| Sauer:beta-Galactosidase Assay (A better Miller)]] | ||
==Proteins: ''in vitro''== | |||
[[Sauer:Silver staining]] | [[Sauer:Silver staining]] | ||
[[Sauer: | [[Sauer:bis-Tris SDS-PAGE, the very best]] | ||
==Nucleic acids: ''in vitro''== | |||
[[Sauer:In vitro transcription with T7 RNA polymerase]] | [[Sauer:In vitro transcription with T7 RNA polymerase]] | ||
| Line 38: | Line 45: | ||
[[Sauer:Phenol/chloroform extraction]] | [[Sauer:Phenol/chloroform extraction]] | ||
[[Sauer: | [[Sauer:Nucleic acid quantitation]] | ||
[[Sauer:Site-directed mutagenesis]] NEEDS TO BE FILLED IN, WORK ON ME! | |||
==More ''in vivo'' goodies== | |||
[[Sauer: | [[Sauer:Competent cell preparation]] NEEDS TO BE FILLED IN, WORK ON ME! | ||
[[ | [[Sauer:P1vir phage transduction]] | ||
Revision as of 13:33, 31 August 2005
Here we will post protocols that people in the lab generally use. Many of these will be lab-specific protocols, but if you think the protocol you are posting is more generally applicable, please link it to the general Protocols page. Also, if you typically do a variation of the protocol that is posted, please list it on the same page as a separate, alternative protocol. If possible, also try to note why what you do is different.
PLEASE NOTE: Blindly following a protocol is not a good substitute for actually talking to someone who has done the experiment. When posting protocols, please also include the name(s) of people in the lab who will be good to talk to about the protocol.
Protein Expression/Purification
Sauer:ClpP purification NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:Expression/purification of 35S-Met proteins
In vitro enzymatic assays
Sauer:In vitro degradation of GFP-ssrA by ClpXP
Sauer:In vitro degradation of radiolabeled proteins by ClpXP NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:ATPase assays NEEDS TO BE FILLED IN, WORK ON ME!
Analysis of proteins or RNA in vivo
Sauer:Western blot NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:Quantitative Western blot NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:Northern blot NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:In vivo tmRNA tagging assays NEEDS TO BE FILLED IN, WORK ON ME!
Sauer:beta-Galactosidase Assay (A better Miller)
Proteins: in vitro
Sauer:bis-Tris SDS-PAGE, the very best
Nucleic acids: in vitro
Sauer:In vitro transcription with T7 RNA polymerase
Sauer:Phenol/chloroform extraction
Sauer:Nucleic acid quantitation
Sauer:Site-directed mutagenesis NEEDS TO BE FILLED IN, WORK ON ME!
More in vivo goodies
Sauer:Competent cell preparation NEEDS TO BE FILLED IN, WORK ON ME!
