Books  Animal & General Biology  Biochemistry & Molecular Biology 

Molecular Biology Techniques: An Intense Laboratory Course

TextbookHandbook / Manual

By: Walt Ream and Katharine G Field

234 pages, no illustrations

Academic Press

Spiralbound | Dec 1999 | #89483 | ISBN: 0125839901
Availability: Usually dispatched within 2-3 weeks Details
NHBS Price: £54.99 $71/€60 approx

About this book

This manual is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern (DNA) blotting, Northern (RNA) blotting, Western blotting, DNA sequencing, oligo-directed mutagenesis, and protein expression. Among its key features, it: provides well-tested experimental protocols for each technique; lists the reagents and preparation of each experiment separately; contains a complete schedule of experiments and the preparation required; and includes study questions at the end of each chapter.


Preface. Course Synopsis: Introduction. Safety Precautions. Daily Schedule. Acknowledgements. Exercises: I. DNA Preparation, Polymerase Chain Reaction, and Molecular Cloning: Cesium chloride-ethidium bromide density gradient centrifugation. PCR to synthesize virD2 flanked with restriction sites. Restriction digests of plasmid pGEX2 and PCR products. Purification of DNA fragments from agarose. Ligation of PCR product to pGEX2 vector. Transformation of E. coli with the ligated plasmid. Small-scale preparation of plasmid DNA by the alkaline lysis method. Restriction analysis. Study questions. Protein Expression, Purification, and Analysis: Expression and purification of a fusion protein. SDS-polyacrylamide gel electrophoresis. Silver stain detection of proteins. Study questions. Oligonucleotide-Directed Mutagenesis: Restriction digests of virD2 (in pCS64) and pUC119. Purification of DNA fragments from agarose. Ligatation of restriction fragment and vector. Transformation of E. coli with the ligated plasmid and recovery clones. Small-scale preparation of plasmid DNA from broth cultures. Restriction digest of DNA: examination to confirm insert. Preparation of single-stranded DNA template. Phosphorylation of oligonucleotide. Annealing mutant oligonuleotide to template. In vitro DNA synthesis by primer extension. Transform synthesis reaction into E. coli DH5a. Small-scale preparation of plasmid DNA. Confirmation of mutants by restriction analysis. Study questions. DNA Sequencing: Polyacrylamide sequencing gel electrophoresis. Didexy sequencing. Automated Sequencing. Introduction to databases and gene sequence analysis. Study questions. Southern Blot Detection of DNA: Preparation of genomic DNA from Agrobacterium tumefaciens. Restriction digestion of genomic DNA. Agarose gel electrophoresis of restriction fragments. Southern blot: denaturation and blotting of DNA gel. Preparation of probe nick translation. Hybridization and washing of Southern blots. Study questions. Northern Blot Detection of mRNA: Preparation of RNA from tobacco leaves. Agarose-formaldehyde gel electrophoresis. Northern blot: denaturation and blotting of RNA. Probe preparation. Hybridization and washing of Northern blots. Study questions. Protein Interaction Analysis in Yeast: Yeast transformation. Filter b-galactosidase assay. Index.

Write a review

There are currently no reviews for this product. Be the first to review this product!

Bestsellers in this subject

Ancestral Sequence Reconstruction

NHBS Price: £32.50 $42/€36 approx

The Cell Cycle

NHBS Price: £33.99 $44/€37 approx

The Ten Most Important Things About Cells

Clearance price: £5.00 £15.00 (Save £10.00) $6/€6 approx

The Cyanobacteria

NHBS Price: £215.00 $277/€236 approx

Molecular Evolution

NHBS Price: £41.99 $54/€46 approx

VAT: GB 407 4846 44
NHBS Ltd is registered in England and Wales: 1875194