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Molecular Biology Protocols

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 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

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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