Technical Protocols

Below is a listing of the protocols that are generally used in the Schimenti Lab. They are extracted from our lab techniques manual, otherwise known as The Joy of Cloning

Some protocols are essentially the same as those described in published lab guides (such as the Maniatis book or the "Red Book"). Others come from other people. Original references are cited in some cases.

Minimal theory, if any, is given. These are cookbook procedures to have at the bench when doing a protocol.

Please contact

if you have any better or faster protocols, as we are always looking to do things easier.

The Joy of Cloning

Table of Contents

Purification of DNA

• Phenol Extraction
• Ethanol Precipitation
• Purification of DNA from Gels
  • Low melting temperature agarose, standard method
  • Agarase Extraction of DNA Fragments
  • Isolation of DNA from Acrylamide Gels

DNA Isolation

• From Tissues
  • Isolation of DNA from Tissues
  • Mouse Tail DNA Prep (the classic)
  • Mouse Brain DNA Prep for Southern Blotting
  • Genotyping Mice by PCR, I
  • Genotyping Mice by PCR,II
  • Genotyping Mice by PCR,III
  • We now use the HotSHOT method...by far the best: Biotechniques 29:52 (2000)
  •  
• From bacteria (plasmids, cosmids)
  • Large Scale Preparation of Plasmid DNA by Alkaline Lysis
  • Mini Plasmid Prep

Recombinant DNA Techniques and Cloning

• Spin Column chromatography
• Southern Blotting
• Oligo labeling of DNA probes
• Colony Lifts and Screening, Standard and Autoclaving Methods

RNA WORK

• acid phenol method
• RNA Formaldehyde for NORTHERN Blotting

ES Cell Work

• Trypsinizing Cells
• DNA Extraction, Standard Method
• Electroporation
• Picking ES cell colonies after electroporation
• Splitting and Freezing
• 96-well Plate Microextraction Protocol
• Genotyping of colonies right after picking

Links

• Schimenti Lab