Sauer:Protocols

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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:In vitro degradation of GFP-ssrA by ClpXP]]
 
[[Sauer:Expression/purification of 35S-Met proteins]]
[[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: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:Site-directed mutagenesis]] NEEDS TO BE FILLED IN, WORK ON ME!
+
[[Sauer:Northern blot]] NEEDS TO BE FILLED IN, WORK ON ME!
-
[[Sauer:Competent cell preparation]] NEEDS TO BE FILLED IN, WORK ON ME!
+
[[Sauer:In vivo tmRNA tagging assays]] NEEDS TO BE FILLED IN, WORK ON ME!
-
[[Sauer:P1vir phage transduction]]
+
[[beta-Galactosidase Assay (A better Miller)| Sauer:beta-Galactosidase Assay (A better Miller)]]
-
[[Sauer:Northern blot]] NEEDS TO BE FILLED IN, WORK ON ME!
+
==Proteins: ''in vitro''==
[[Sauer:Silver staining]]
[[Sauer:Silver staining]]
-
[[Sauer:In vivo tmRNA tagging assays]] NEEDS TO BE FILLED IN, WORK ON ME!
+
[[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]]
[[Sauer:Phenol/chloroform extraction]]
[[Sauer:Phenol/chloroform extraction]]
-
 
-
[[Sauer:bis-Tris SDS-PAGE, the very best]]
 
[[Sauer:Nucleic acid quantitation]]
[[Sauer:Nucleic acid quantitation]]
-
[[beta-Galactosidase Assay (A better Miller)| Sauer:beta-Galactosidase Assay (A better Miller)]]
+
[[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!
 +
 
 +
[[Sauer:P1vir phage transduction]]

Revision as of 15:33, 31 August 2005

SAUER LAB

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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.

Contents

Protein Expression/Purification

Sauer:ClpX 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:Silver staining

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!

Sauer:P1vir phage transduction

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