Knight:Protocols

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(Protein)
Current revision (19:46, 29 November 2009) (view source)
(DNA)
 
(25 intermediate revisions not shown.)
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==DNA==
==DNA==
[[Knight:Agarose gel electrophoresis]]
[[Knight:Agarose gel electrophoresis]]
 +
 +
[[Knight:Annealing and cloning oligos]] -- <font color=red>in progress, hasn't been tested</font>
[[Annealing and primer extension]]
[[Annealing and primer extension]]
 +
 +
[[Knight:Annealing and primer extension with Klenow polymerase]]
[[Knight:Annealing and primer extension with Taq polymerase]]
[[Knight:Annealing and primer extension with Taq polymerase]]
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[[Knight:DNA ligation using NEB Quick Ligation Kit]]
[[Knight:DNA ligation using NEB Quick Ligation Kit]]
 +
 +
[[Knight:DNA ligation using T4 DNA ligase]]
 +
 +
[[Knight:Maxiprep]]
[[Knight:Micropure EZ and Microcon purification]]
[[Knight:Micropure EZ and Microcon purification]]
[[Miniprep]]
[[Miniprep]]
 +
 +
[[Knight:Miniprep low copy plasmids]]
[[Phosphatase treatment of linearized vector]]
[[Phosphatase treatment of linearized vector]]
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[[Knight:Sequencing DNA]]
[[Knight:Sequencing DNA]]
 +
 +
[[Knight:TempliPhi]]
[[Knight:TOPO TA cloning]]
[[Knight:TOPO TA cloning]]
 +
 +
[[Knight:TOPO vector preparation]]
[[Knight:Addition of 3' A-overhangs]]
[[Knight:Addition of 3' A-overhangs]]
[[Knight:Site-directed mutagenesis]]
[[Knight:Site-directed mutagenesis]]
 +
 +
[[Norville:Test site]]
 +
 +
==RNA==
 +
[[Knight:In vitro transcription]]
 +
 +
[[Knight:RNA electrophoresis]]
==Protein==
==Protein==
 +
[[Knight:Protein solubility]]
 +
[[Knight:Centrifuge desalting]]
[[Knight:Centrifuge desalting]]
-
[[Knight:Dialysis]]
+
[[Knight:Dialysis]] --<font color=red>works poorly</font>
-
[[Knight:Electrophoretic mobility shift assay]] --<font color=red>in progress!</font>
+
[[Knight:Electrophoretic mobility shift assay]]
-
[[Knight:Gel filtration]] --<font color=red>in progress!</font>
+
[[Knight:Gel filtration]]
-
[[Knight:Microcon filtration of proteins]]
+
[[Knight:Microcon filtration of proteins]] --<font color=red>works poorly</font>
[[Knight:NuPAGE electrophoresis]]  
[[Knight:NuPAGE electrophoresis]]  
-
[[Knight:Protein DNA binding]] --<font color=red>in progress!</font>
+
[[Knight:Protein DNA binding]]
[[Knight:Purification of His-tagged proteins]]
[[Knight:Purification of His-tagged proteins]]
 +
 +
[[Knight:Purification of MBP-tagged proteins]]
==''Escherichia coli''==
==''Escherichia coli''==
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[[Transforming chemically competent cells (Inoue)]]
[[Transforming chemically competent cells (Inoue)]]
 +
 +
[[TOP10 chemically competent cells]] - preferred by TK and the Registry
[[Knight:Preparing electrocompetent cells]]
[[Knight:Preparing electrocompetent cells]]
[[Knight:Electroporation]]
[[Knight:Electroporation]]
 +
 +
[[Knight:DNA Spots]]
 +
 +
[[Knight:Beta-galactosidase assay]]
 +
 +
[[Knight:Beta-galactosidase assay/96 well format]]
 +
 +
[[Knight:Preparing stabs]]
==Miscellaneous==
==Miscellaneous==
 +
[[Knight:Deionization]]
 +
 +
[[Knight:Dishwashing]]
 +
[[Knight:pH meter/Calibration|Calibrating the pH meter]]
[[Knight:pH meter/Calibration|Calibrating the pH meter]]

Current revision

CSAIL logo
Knight Lab



Contents

DNA

Knight:Agarose gel electrophoresis

Knight:Annealing and cloning oligos -- in progress, hasn't been tested

Annealing and primer extension

Knight:Annealing and primer extension with Klenow polymerase

Knight:Annealing and primer extension with Taq polymerase

Knight:Colony PCR

Ethanol precipitation of small DNA fragments

Knight:DNA ligation using NEB Quick Ligation Kit

Knight:DNA ligation using T4 DNA ligase

Knight:Maxiprep

Knight:Micropure EZ and Microcon purification

Miniprep

Knight:Miniprep low copy plasmids

Phosphatase treatment of linearized vector

Knight:Reconstituting primers

Knight:Restriction Digest

Knight:Sequencing DNA

Knight:TempliPhi

Knight:TOPO TA cloning

Knight:TOPO vector preparation

Knight:Addition of 3' A-overhangs

Knight:Site-directed mutagenesis

Norville:Test site

RNA

Knight:In vitro transcription

Knight:RNA electrophoresis

Protein

Knight:Protein solubility

Knight:Centrifuge desalting

Knight:Dialysis --works poorly

Knight:Electrophoretic mobility shift assay

Knight:Gel filtration

Knight:Microcon filtration of proteins --works poorly

Knight:NuPAGE electrophoresis

Knight:Protein DNA binding

Knight:Purification of His-tagged proteins

Knight:Purification of MBP-tagged proteins

Escherichia coli

Preparing chemically competent cells (Inoue)

Transforming chemically competent cells (Inoue)

TOP10 chemically competent cells - preferred by TK and the Registry

Knight:Preparing electrocompetent cells

Knight:Electroporation

Knight:DNA Spots

Knight:Beta-galactosidase assay

Knight:Beta-galactosidase assay/96 well format

Knight:Preparing stabs

Miscellaneous

Knight:Deionization

Knight:Dishwashing

Calibrating the pH meter

Measuring pH

Knight:Poster printing

Personal tools