CPC Molecular Biology Protocols

This resource attempts to offer a comprehensive collection of molecular biology and Nematode-related protocols. The site links to protocols either maintained directly by the Ambros Lab or at other locations around the world.

RNA
RNA Preparation and Treatment
Northern Blotting: Efficient RNA Staining and Transfer: Kim Marshall
Northern Blots: Michael Koelle
RNA Preparation: MGH
LiCl RNA Preparation: MGH
Worm RNA Preparation: MGH
C. elegans RNA prep: Michael Koelle
DNA
DNA Transformation/Library Preparation NMFS/NOAA
Transformation-Competent E. coli preparation (rubidium chloride method)
Cosmid packaging protocol


Plasmid DNA
Genome Sequencing Center Protocol List
Plasmid min iprepI: GSC
Plasmid miniprepII: GSC
Mini-Prep Procedure For E. coli Plasmid Purification: Kim Marshall
Low copy plasmid purification using Qiagen Midi columns: NMFS/NOAA
Large Scale Plasmid Preparation: MGH
DNA purification using Diatomaceous Earth, (plasmids, cosmids and p1): NMFS/NOAA
Introduction
Maxi-preps and all media, solutions
Midi-preps
Mini-preps


Cosmid DNA
Codes for cosmid vectors: GSC
Cosmid fingerprinting: GSC


Phage DNA
PCR of M13 inserts: GSC
Preparation of M13 sublibraries: GSC
M13 RF prep largescale: GSC
M13 ThermoMAX prep: GSC
M13 helperphage: GSC
M13 PEG prepI: GSC
M13 PEG prepII: GSC
Largescale M13 prep: GSC
Lambda DNA Miniprep: Kim Marshall
Single stranded template from pBluescript phagemid: NMFS/NOAA


Genomic DNA
Molecular Cytogenetic Genome ScreenComparative Genomic Hybridization
Metaphase chromosome preparation
DNA preparation by cryostom tissue dissection
DNA labeling by nick translation
Hybridization
DNA detection
Image capture
Image analysis
DAPI chromosome identification
Southern Blots
Worm genomic Southern blots: Michael Koelle
Alkaline Southern Blotting Procedure: Kim Marshall
DNA purification, chromosomal, triton extraction method: NMFS/NOAA
DNA purification, bacterial-genomic via Qiagen columns: NMFS/NOAA
Worm Genomic DNA: MGH
YACs
Easy YAC Preparation MethodDavies


Fragments
DNA isolation from low melting-temperature agarose: NMFS/NOAA


DNA Sequencing
Direct Automated Sequencing off Bacterial Genomic DNABruce Rhoe
Sequencing through stop regions of M13 or pUC subclones, and PCR productsGSC
Terminator reaction clean-up: GSC
Linear amp seqI: GSC
Linear amp seqII: GSC
Linear amp seqIII: GSC
Linear amp seqIV Licor: GSC
Sequencing gel solutions: NMFS/NOAA
Sequencing gels: wedge gel performance without pouring wedge gels: NMFS/NOAA
PCR and Related Methods
Degenerate PCR Protocol: M. Koelle
Long PCR Protocol: Church Lab
Tc1 Deletion PCR Tips: MGH
Core sample PCR: a method to re-PCR unique bands from products of mixed size: NMFS/NOAA
Cloning PCR-Generated DNA Fragments: NMFS/NOAA, submitted by Hugh Griffin, 7/7/95
Site-directed Mutagenesis using PCR: NMFS/NOAA, submitted by Hugh Griffin, 7/25/95
Proteins
Protein
Protein Electrophoresis
Tricine-polyacrylamide gels for high resolution of small proteins: NMFS/NOAA
Protein & Peptide HPLC Columns
HILIC and SEC Columns PolySULFOETHYL and PolyHYDROXYETHYL Columns
Polyaspartic Acid?Weak Cation Exchange Columns
PolySULFOETHYL Aspartamide?Strong Cation Exchange Columns
(ALKYL) Aspartamide? HIC Columns
PolyWAX LP? Weak Anion Exchange Columns
PolyGLYCOPLEX? Complex Carbohydrates Columns
SDS Removal Guard Columns
PolyLC Part Numbers and Prices
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