Knight:Protocols: Difference between revisions
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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: | [[Knight:Protein solubility]] | ||
[[Knight: | [[Knight:Centrifuge desalting]] | ||
[[Knight:Microcon filtration of proteins]] | [[Knight:Dialysis]] --<font color=red>works poorly</font> | ||
[[Knight:Electrophoretic mobility shift assay]] | |||
[[Knight:Gel filtration]] | |||
[[Knight:Microcon filtration of proteins]] --<font color=red>works poorly</font> | |||
[[Knight:NuPAGE electrophoresis]] | [[Knight:NuPAGE electrophoresis]] | ||
[[Knight:Protein DNA binding]] | [[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]] | ||
Latest revision as of 16:46, 29 November 2009
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Knight Lab
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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
Ethanol precipitation of small DNA fragments
Knight:DNA ligation using NEB Quick Ligation Kit
Knight:DNA ligation using T4 DNA ligase
Knight:Micropure EZ and Microcon purification
Knight:Miniprep low copy plasmids
Phosphatase treatment of linearized vector
Knight:TOPO vector preparation
Knight:Addition of 3' A-overhangs
Knight:Site-directed mutagenesis
RNA
Protein
Knight:Dialysis --works poorly
Knight:Electrophoretic mobility shift assay
Knight:Microcon filtration of proteins --works poorly
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:Beta-galactosidase assay
Knight:Beta-galactosidase assay/96 well format
