Manipulations of E. coli

 

E. coli strains

 

Growth media

 

Transformation of E. coli

 

 

Transformation by the calcium chloride procedure (modified from Mandel and Higa, )

 

 

Transformation by electroporation (Dower et al. )

 

 

Transformation of E. coli (Ausubel et al) - quick protocol

 

 

Preparation of electrocompetente cells (volgens Tung & Chow)

 

 

Transformation- and mutagenesis-protocol met Epicurian Coli XL-Red

 

 

Transformation of E. coli (according to Inouoe et al )

 

Phase contrast microscopy

 

Determination of minimal inhibitory concentration (MIC) of chloramphenicol

 

 

In vivo karakterisatie van CatI-mutanten

Yeast manipulations

 

Saccharomyces cerevisiae strains

 

Growth media for Saccharomyces cerevisiae

 

Determination of cell density

 

Transformation of S. cerevisiae

 

 

Transformation by the lithium acetate (LiOAc) method (Ito et al, ; Gietz and Shiestl, )

 

 

Electroporation of Yeast

 

 

High efficiency method for LiAc/SS-DNA/PEG transformation of yeast

 

 

Quick and Easy TRAFO Protocol for yeast transformation

 

 

Direct transfer of plasmid DNA from yeast to E. coli by electroporation

 

Preparation of protein extracts from yeast

 

Sporulation of yeast

 

 

Protocol 1: Sporulation in liquid medium (Kassir & Simchen, )

 

 

Protocol 2: Sporulation on solid medium (Kassir & Simchen, )

 

 

Protocol 3: Sporulation in liquid medium (Codón et al, )

 

Preparation of mass spores of yeast

 

Tetrad disection of yeast

DNA manipulations

 

DNA preparations

 

 

Mini Plasmid preparation from E. coli cultures (Serghini et al, )

 

 

Qiagen Midi (tip ) plasmid preparation from E. coli cultures (Qiagen GmbH, Düsseldorf, Germany)

 

 

QIAprep Miniprep plasmid purification from E. coli cultures

 

 

Fast small scale isolation of yeast total DNA (Method of Wach et al.)

 

 

Fast small scale isolation of total yeast DNA (Hoffman and Winston, )

 

DNA concentration determination

 

Enzymatic DNA manipulations

 

 

Purification after ligation

 

Oligonucleotide synthesis

 

Polymerase chain reaction (PCR)

 

 

Normal PCR

 

 

Inverse PCR

 

 

Randommutagenesis using PCR

 

PCR clone analysis on intact E. coli cells

 

DNA sequence analysis

 

Preparation of Sephadex-G

Protein manipulations

 

Protein purification

 

 

Dialysis of CAT-proteins

 

 

Pefablock

 

Protein concentration determination

 

 

Total protein concentration

 

 

Determination of CAT concentration

 

SDS polyacrylamide gel electrophoresis (SDS-PAGE) analysis

 

Determination of protein solubility

 

CAT activity assay

 

Determination of thermal inactivation kinetics

 

Fourier transform infrared (FT-IR) analysis

 

Protocol CAT-ELISA (Boehringer Manheim Biochemica)

 

Overzicht van protocols en ideeën om de denaturatie/renaturatie van eiwitten efficiënter te laten verlopen

 

Isolatieprocedure inclusiepartikels hervouwing van eiwitten uit IB's

Software

 

Hoe PDB-coördinaten omzetten in data bruikbaar voor Nanovision ?