Paulsson:Protocols

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Many excellent protocols can be found in Current Protocols in Molecular Biology, available through the Countway Digital Library.

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Preparation of electrocompetent E.coli
Electroporation of E.coli
Measuring DNA replication with 3H-thymidine

Nanodrop
Plate Reader

Getting Started In the Lab

Pipetting 101

general
what volume, which tool
accuracy and calibration (cleaning)

Sterile technique

Nurturing Microbes

preservation growth conditions genotypes antibiotics

Media Preparation

Current Protocols in Molecular Biology article on mimimal, rich, solid and agar media for E. coli
Media:Ecoli_preparation_media.pdf

  • MSC media (from Rosenfeld et al., Science v307, 25 March 2005, SOM)

M9 salts
0.6% succinate
0.01% casamino acids
0.15 ug/ml biotin
1.5 uM thiamine
doubling time for E. coli reported to be 45 (-/+10) min at 32 degrees
for microscope pad, dissolve low melt agarose in MSC (1.5%)

Plasmid Biology

definitions transformation isolation segragation incompatability copy number selection


Manipulating DNA

Synthesis PCR Enzymes
Quick Change mutagenesis

Agarose Gels

For small casting tray - 60 ml volume