CGH Protocols
Metaphase chromosome preparation
DNA preparation by cryostom tissue dissection
DNA labeling by nick translation
Hybridization
DNA detection
Image capture
Image analysis
DAPI chromosome identification
Pulsed-field Gel Purification of E. coli Restriction Fragments
Sequencing Library Construction
Salk Flow Cytometry Protocols
Cell cycle - propidium iodide only
FCM of leukocytes in whole blood
Table of Contents
A. Phenol extraction of DNA samples
B. Concentration of DNA by ethanol precipitation
D. Agarose gel electrophoresis
E. Elution of DNA fragments from agarose
H. Fragment purification on Sephacryl S-500 spin columns
II. Random subclone generation
C. Random fragment end-repair, size selection, and phosphorylation
F. Bacterial cell transformation
G. Microcentrifuge tube transformation
III. Methods for DNA isolation
A. Large scale double-stranded DNA isolation
B. Midiprep double-stranded DNA isolation
C. Miniprep double-stranded DNA isolation
D. Large scale M13RF isolation
E. Single-stranded M13 DNA isolation using phenol
F. Biomek-automated modified-Eperon isolation procedure for single-stranded M13DNA
G. 96 well double-stranded template isolation
H. Genomic DNA isolation from blood
IV. Methods for DNA sequencing
A. Bst-catalyzed radiolabeled DNA sequencing
B. Radiolabeled sequencing gel preparation, loading, and electrophoresis
C. Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers
D. Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye terminator reactions
1. Terminator Reaction Clean-Up via Centri-Sep Columns
2. Terminator Reaction Clean-Up via Sephadex G-50 Filled Microtiter Format Filter Plates
E. Sequenase[TM] catalyzed sequencing with dye-labeled terminators
H. cDNA sequencing based on PCR and random shotgun cloning
A. Polymerase Chain Reaction (PCR)
B. Purification of PCR fragments for cloning
D. Synthesis and purification of oligonucleotides
E. Rapid hybridization of complementary M13 inserts
Taq Cycle Sequencing Reagent Preparation
Oligonucleotide universal primers used for DNA sequencing
Listing of M13 (pUC) cloning sites
Commonly used restriction enzymes and assay buffers
Bacterial Transformation and Transfection
Codon chart and amino acid symbols
Biomek configuration for single stranded DNA isolation
Molecular Biology Laboratory Manual
CHAPTER 1: General Laboratory Methods
CHAPTER 2: Instructions for Notebook Keeping
CHAPTER 3: Vector NTI User's Guide
CHAPTER 4: Molecular Biology Methods
Preparation of genomic DNA from bacteria
Restriction enzyme digestion of DNA
Phenol/chloroform extraction of DNA
Transformation of E. coli by electroporation
Preparative DNA Fragment Isolation from an Agarose Gel
Ligations of plasmid DNA to insert DNA
Transfection of mammalian cells using Lipofectamine
Western Blot analysis of His-tagged proteinsl
Cycle Sequencing Protocols For ABI 3100
One Step Gene Assembly (Gene Synthesis)
CHAPTER 5: Tissue Culture Methods
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