Technique / Molecular Biology


Molecular Biology Protocols



Molecular Biology Protocols

NWFSC Fish Health/Microbiology Molecular Biology Protocols (served since 1995).

DNA Purification Techniques

DNA purification (plasmids, cosmids and p1) using Diatomaceous Earth (Bruce Roe Lab)

Introduction
Maxi-preps and all media, solutions
Midi-preps
Mini-preps

DNA purification, plasmid mini-preps, diatomaceous earth, contributed by Kyung-Soo Kim and Charles K. Pallaghy 2/10/97
DNA purification, bacterial-genomic, triton extraction method
DNA purification, bacterial-genomic via Qiagen columns
DNA purification, bacterial genomic, specifically Gram positive bacteria
DNA isolation from low melting-temperature agarose
Low copy plasmid purification using Qiagen Midi columns
Plasmid "Miniprep" Isolation: tried and true!
Single stranded template from pBluescript phagemid

DNA Transformation/Library Preparation

Transformation-Competent E. coli preparation

Rubidium chloride method
Inoue "ultra-competent" method

Cosmid packaging protocol


DNA Ligation and Transformation Protocols (PDF format)
(contributed by Tim Fitzwater 8/15/02)

Southern/Northern Blotting

Southern blot-chemiluminescent detection, using Lumigen or Lumiphos (BM "Genius" system)
Southern blot-chemiluminescent detection, using CSPD (BM "Genius" system)

DNA Sequencing

Sequencing gel solutions
Sequencing gels: wedge gel performance without pouring wedge gels

Oligonucleotides

Oligonucleotide Visualization
Oligonucleotide calculator: determine Tm, MW, etc. of your oligonucleotide (requires javascript enabled browser)

PCR and Related Methods

Core sample PCR: a method to re-PCR unique bands from products of mixed size
Cloning PCR-Generated DNA Fragments, contributed by Hugh Griffin, 7/7/95
Site-directed Mutagenesis using PCR, contributed by Hugh Griffin, 7/25/95
Reverse Transcriptase PCR, contributed by Jim Hutchins 8/25/95
Error Rates for Thermal Resistant DNA Polymerases
Red Dye, Taq Polymerase and PCR, contributed by Tim Fitzwater 2/10/03.

RNA Methods

Solublization of RNA in Formamide, contributed by James McCaughern-Carucci, 12/31/96
RNAse Protection Assay, contributed by James McCaughern-Carucci, 1/9/97
The "Neverfail" Northern Blot Hybridization, contributed by James McCaughern-Carucci, 10/26/97
Northern blots, small RNA protocol, contributed by Tim Fitzwater, 3/20/03

Protein Electrophoresis

Tricine-polyacrylamide gels: for high resolution of small proteins
SDS-polyacrylamide gels: standard Laemmli protocol
Ethanol-based SDS-PAGE Coomassie Blue staining, contributed by Roger Rowlett, 9/5/97
Bench protocol for silver staining SDS-polyacrylamide gels, contributed by Melissa Caimano, 3/20/00
SDS-PAGE troubleshooting: David R. Caprette's humorous yet extremely informative site on how to troubleshoot problems with SDS-PAGE gels (opens off-site link in a new window)
Protein Purification
Preparation of active proteins from inclusion bodies, contributed by Laurent Vuillard and Alasdair Freeman, 3/11/99
Autoradiography Tips
Intensifying Screens -- their use explained, contributed by John Attwood 12/1/97
Safety
Generation of Phosgene from Chloroform !
Other References

Standard Genetic Code
Links to other Molecular Biology Protocols and Resources

Last update 28-Nov-2004, Rating of 0 votes.

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