Sauer:Protocols

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(Protein Expression/Purification)
 
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{{Sauer lab sidebar}}
{{Sauer lab sidebar}}
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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.
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Please link protocols to the general [[Protocols]] page if appropriate.
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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.
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<div style="padding: .4em .9em .9em">
<div style="padding: .4em .9em .9em">
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==Protein Expression/Purification==
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===Protein Expression/Purification===
[[Sauer:ClpX purification]]
[[Sauer:ClpX purification]]
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[[Sauer:ClpP purification]] PROTOCOL NEEDED!
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[[Sauer:His6 ClpX purification]]
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[[Sauer:ClpP purification]]  
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[[Sauer:ClpA purification]]
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[[Sauer:SspB purification]]
[[Sauer:Expression/purification of 35S-Met proteins]]
[[Sauer:Expression/purification of 35S-Met proteins]]
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==''In vitro'' enzymatic assays==
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[[Sauer:Purification of His-tagged proteins|Sauer:Purification of His<sub>6</sub>-tagged proteins]]
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[[Sauer:Lysing E. coli with Lysozymes]]
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[[Sauer:GFP-H6-ssrA purification]]
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[[Sauer:GST-sFtsH purification]]
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[[Sauer:FtsH purification]]
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===''In vitro'' enzymatic assays===
[[Sauer:In vitro degradation of GFP-ssrA by ClpXP]]
[[Sauer:In vitro degradation of GFP-ssrA by ClpXP]]
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[[Sauer:In vitro degradation of GFP-ssrA by ClpAP]]
[[Sauer:In vitro degradation of radiolabeled proteins by ClpXP]] PROTOCOL NEEDED!
[[Sauer:In vitro degradation of radiolabeled proteins by ClpXP]] PROTOCOL NEEDED!
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[[Sauer:ATPase assays]] PROTOCOL NEEDED!
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[[Sauer:ATPase assays]]
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==Proteins: ''in vitro''==
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[[Sauer:In vitro peptide degradation by ClpXP]]
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===Proteins: ''in vitro''===
[[Sauer:Silver staining]]
[[Sauer:Silver staining]]
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[[Sauer:bis-Tris SDS-PAGE, the very best]]
[[Sauer:bis-Tris SDS-PAGE, the very best]]
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==Nucleic acids: ''in vitro''==
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===Nucleic acids: ''in vitro''===
[[Sauer:In vitro transcription with T7 RNA polymerase]]
[[Sauer:In vitro transcription with T7 RNA polymerase]]
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[[Sauer:Phenol/chloroform extraction]]
[[Sauer:Phenol/chloroform extraction]]
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[[Sauer:Nucleic acid quantitation]]
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[[Quantitating nucleic acids]]
[[Sauer:Site-directed mutagenesis]]
[[Sauer:Site-directed mutagenesis]]
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[[Sauer:RNA Purification from E. coli]]
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[[Sauer:Stitching Genes by PCR]]
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<div style="clear: right; text-align: left; float: left; padding: .4em .9em .9em">
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==Analysis of proteins or RNA ''in vivo''==
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===Analysis of proteins or RNA ''in vivo''===
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[[Sauer:Western blot]] PROTOCOL NEEDED!
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[[Sauer:Western blot]]
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[[Sauer:Quantitative Western blot]] PROTOCOL NEEDED!
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[[Sauer:Quantitative Western blot]]
[[Sauer:Northern blot]] PROTOCOL NEEDED!
[[Sauer:Northern blot]] PROTOCOL NEEDED!
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[[beta-Galactosidase Assay (A better Miller)| Sauer:beta-Galactosidase Assay (A better Miller)]]
[[beta-Galactosidase Assay (A better Miller)| Sauer:beta-Galactosidase Assay (A better Miller)]]
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==More ''in vivo'' goodies==
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===More ''in vivo'' goodies===
[[Sauer:Competent cell preparation]] PROTOCOL NEEDED!
[[Sauer:Competent cell preparation]] PROTOCOL NEEDED!
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[[Sauer:P1vir phage transduction]]
[[Sauer:P1vir phage transduction]]
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==Other good stuff==
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[[Sauer:Plasmid-born λRed recombineering using dsDNA]]
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===Other good stuff===
[[Library Generation|Generation of genomic libraries]]
[[Library Generation|Generation of genomic libraries]]
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[[Sauer:HPLC protocol]]
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[[Sauer:FPLC protocol]]
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Current revision

SAUER LAB

Home        Protocols        Lab Members        Materials        Equipment        Links        Internal       

Please link protocols to the general Protocols page if appropriate.

Protein Expression/Purification

Sauer:ClpX purification

Sauer:His6 ClpX purification

Sauer:ClpP purification

Sauer:ClpA purification

Sauer:SspB purification

Sauer:Expression/purification of 35S-Met proteins

Sauer:Purification of His6-tagged proteins

Sauer:Lysing E. coli with Lysozymes

Sauer:GFP-H6-ssrA purification

Sauer:GST-sFtsH purification

Sauer:FtsH purification

In vitro enzymatic assays

Sauer:In vitro degradation of GFP-ssrA by ClpXP

Sauer:In vitro degradation of GFP-ssrA by ClpAP

Sauer:In vitro degradation of radiolabeled proteins by ClpXP PROTOCOL NEEDED!

Sauer:ATPase assays

Sauer:In vitro peptide degradation by ClpXP

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

Quantitating nucleic acids

Sauer:Site-directed mutagenesis

Sauer:RNA Purification from E. coli

Sauer:Stitching Genes by PCR



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