Nucleic Acids / edited by John M. Walker.

Format:

Book

Contributor:

Walker, John M., 1948- editor.

SpringerLink (Online service)

Series:

Methods in molecular biology 1064-3745 ; 2.

Springer Protocols (Springer-12345)

Methods in Molecular Biology, 1064-3745 ; 2

Language:

English

Subjects (All):

Life sciences.

Biochemistry.

Life Sciences.

Biochemistry, general.

Local Subjects:

Life Sciences.

Biochemistry, general.

Physical Description:

1 online resource (XIV, 375 pages).

Contained In:

Springer eBooks

Place of Publication:

Totowa, NJ : Humana Press, 1984.

System Details:

text file PDF

Summary:

In recent years there has been a tremendous increase in our understanding of the functioning of the cell at the molecular level. This has been achieved in the main by the invention and development of new methodology, parti- larly in that area generally referred to as "genetic en- neering". While this revolution has been taking place in the field of nucleic acids research, the protein chemist has at the same time developed fresh methodology to keep pace with the requirements of present day molecular bi- ogy. Today's molecular biologist can no longer be content with being an expert in one particular area alone. He/she needs to be equally competent in the laboratory at h- dling DNA, RNA, and proteins, moving from one area to another as required by the problem he/she is trying to solve. Although many of the new techniques in molecular biology are relatively easy to master, it is often difficult for a researcher to obtain all the relevant information nec- sary for setting up and successfully applying a new te- nique. Information is of course available in the research l- erature, but this often lacks the depth of description that the new user requires. This requirement for in-depth pr- tical details has become apparent by the considerable - mand for places on our Molecular Biology Workshops held at Hatfield each summer.

Contents:

The Burton Assay for DNA

DABA Fluorescence Assay for Submicrogram Amounts of DNA

Preparation of "RNase-Free" DNase by Alkylation

The Isolation of Satellite DNA by Density Gradient Centrifugation

The Isolation of High Molecular Weight Eukaryotic DNA

Preparation of Lyophilized Cells to Preserve Enzyme Activities and High Molecular Weight Nucleic Acids

Agarose Gel Electrophoresis of DNA

Autoradiography of Gels Containing 32P

Detection of Specific DNA Sequences-The Southern Transfer

The Extraction and Isolation of DNA from Gels

One-Dimensional Electrophoresis of Nucleic Acids in Agarose Using Denaturation with Formaldehyde and Identification of 3H-Labeled RNA by Fluorography

Gel Electrophoresis of RNA in Agarose and Polyacrylamide Under Nondenaturing Conditions

The Extraction of Total RNA by the Detergent and Phenol Method

RNA Extraction by the Proteinase K Method

RNA Extraction by the Guanidine Thiocyanate Procedure

The Purification of Poly(A)-Containing RNA by Affinity Chromatography

Messenger RNA Fractionation on Neutral Sucrose Gradients

The Estimation of mRNA Content by Poly(U) Hybridization

DNA Directed in Vitro Protein Synthesis with Escherichia coli S-30 Extracts

In Vitro Translation of Messenger RNA in a Wheat Germ Extract Cell-Free System

In Vitro Translation of Messenger RNA in a Rabbit Reticulocyte Lysate Cell-Free System

Immunoprecipitation of in Vitro Translation Products with Protein A Bound to Sepharose

In Vitro Continuation of RNA Synthesis Initiated in Vivo

Synthesis of Double-Stranded Complementary DNA from Poly(A)+mRNA

Plasmid DNA Isolation by the Cleared Lysate Method

Plasmid DNA Isolation (Sheared Lysate Method)

Small-Scale Plasmid DNA Preparation

Preparation of Chromosomal DNA from E. coli

Preparation and Assay of Phage Lambda

Preparation of Phage Lambda DNA

The Use of Restriction Endonucleases

Ligation of DNA with T4 DNA Ligase

The Use of Alkaline Phosphatase to Prevent Vector Regeneration

Bacterial Transformation

Bacterial Transformation (Kushner Method)

In Vitro Packaging of DNA

Yeast Transformation

Radiolabeling of DNA by Nick Translation

Radiolabeling of DNA Using Polynucleotide Kinase

Radiolabeling of DNA with 3? Terminal Transferase

Radiolabeling of DNA with the Klenow Fragment of DNA Polymerase

Identification of Recombinant Plasmids by In Situ Colony Hybridization

Identification of Recombinant Phages by Plaque Hybridization

Plasmid Screening Using Single Colony Lysates

Immunological Detection of Gene Expression in Recombinant Clones

The Isolation of Minicells

The Isolation of Maxicells

The Identification of Gene Products in Minicells and Maxicells

Molecular Cloning in Bacteriophage Lambda and in Cosmids

DNA Transformation of Mammalian Cells

Chemical Cleavage (Maxam and Gilbert) Method for DNA Sequence Determination

Gel Electrophoretic Analysis of DNA Sequencing Products

DNA Sequence Determination Using Dideoxy Analogs.

Other Format:

Printed edition:

ISBN:

9781592594894

Access Restriction:

Restricted for use by site license.