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

Shipping Information
Nanodrop
Plate Reader

Pipetting 101

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

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

Cloning Synthetic Oligonucleotide Cassettes
Quick Change mutagenesis
Colony PCR

Agarose Gels

• For small casting tray, 0.6g agarose + 60 ml TAE, microwave 50-60 sec, add 3ul EthBr stock, let stand for 7 min before pouring
  

Narrow wells (15 lanes): 1.5 x 2.59mm, ~19ul volume
Wide wells (8 lanes): 1.5 x 5.54mm, ~40ul volume

• For large casting tray, 150ml volume, microwave 80-90 sec, add 7.5ul EthBr stock, let stand 10min before pouring
  

Narrow wells (20 lanes): 1.5 x ##mm, ~##ul volume
Wide wells ( lanes): 1.5 x ##mm, ~##ul volume
Multi-channel pipet compatible wells (26 lanes): 1.5 x mm, ~##ul volume