FRONTIERS IN BIOSCIENCE;
PROTOCOLS



[Molecular biology protocols]
[Machines, Media and Reagents]
[Purification]
[Manipulation]
[Transformation/Library Preparation]
[Blotting]
[Sequencing]
[Oligonucleotides]
[PCR]
[Protein Electrophoresis]
C elegans protocols
Molecular biology protocols (1)
Molecular biology protocols (2)
Other biology protocols

Machines, Media and Reagents

ABI 373 Loading Guide
ABI 377 Loading Guide
Media recipes
Molecular reagent list
Plate Pouring

DNA Purification

[Plasmids] [Phage(mids)] [Cosmids] [Diatomaceous Earth] [Special cases] [Reprecipitation]
Plasmids

Plasmid mini-prep I
Plasmid mini-prep II

Phages/Phagemids

Large scale M13 prep
M13 0.5 Thermo MAX prep
M13 PEG prep I
M13 PEG prep II
M13 RF prep large scale
M13 Thermo MAX prep
M13 helper phage
Single stranded template from pBluescript phagemid

Cosmids

Cosmid prep
Codes for cosmid vectors
Cosmid 96 well mini-prep
Cosmid finger printing

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

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

Special cases

DNA isolation from low melting-temperature agarose
DNA purification, bacterial-genomic, triton extraction method
DNA purification, bacterial-genomic via Qiagen columns
Low copy plasmid purification using Qiagen Midi columns

Reprecipitation

DNA/RNA precipitation

DNA Manipulation

PUC pooling
Restriction Digest
T7 gene 6 exo
TA Cloning
Vector preparation
Vectors

DNA Transformation/Library Preparation

Transformation-Competent E. coli preparation

CaCl Transformation
Electrotransformation
Inoue "ultra-competent" method
JS5 Transformation
Rubidium chloride method

Cosmid and phagemid preparation

Cosmid packaging protocol
Preparation of M13 sub-libraries

Southern/Northern Blotting

Southern blot-chemiluminescent detection (BM "Genius" system)

DNA Sequencing

AGTC 96 well mini-prep
Sequencing gel solutions
Sequencing gels: wedge gel performance without pouring wedge gels
Sequenase 1 dye-termination
Sequenase II ALF primers
Sequenase V2.0 licor
Terminator reaction cleanup

Oligonucleotides

Oligonucleotide Visualization
Oligo nucleotides

PCR and Related Methods

Core sample PCR: a method to re-PCR unique bands from products of mixed size
Cloning PCR-Generated DNA Fragments, submitted by Hugh Griffin, 7/7/95
Site-directed Mutagenesis using PCR, submitted by Hugh Griffin, 7/25/95
Reverse Transcriptase PCR, submitted by Jim Hutchins 8/25/95
Error Rates for Thermal Resistant DNA Polymerases
Linear amplification sequencing I
Linear amplification sequencing II
Linear amplification sequencing III
Linear amplification sequencing IV Licor
PCR of M13 inserts
ss PCR magnetic beads

Protein Electrophoresis

Tricine-polyacrylamide gels: for high resolution of small proteins
SDS-polyacrylamide gels: standard Laemmli protocol