Knight:Protocols

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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