| Description |
1 online resource |
| Contents |
Part I. Manipulation of DNA -- Getting oriented: practicing with micropipettes -- Purification and digestion of plasmid (vector) DNA -- Completion of vector preparation and polymerase chain reaction amplification of egfp -- Preparation on insert DNA (egfp) PCR product -- DNA ligation and transformation of Escherichia coli -- Part II. Screening transformants -- Screening of transformants, part I -- Screening of transformants, part II -- Analysis of DNA sequence from a positive clone -- Part III. Expression, detection, and purification of recombinant proteins from bacteria -- Expressions of fusion protein from positive clones, SDS-PAGE and Western Blot: Part I -- Expressions of fusion protein from positive clones, SDS-PAGE and Western Blot: Part II -- Extraction of recombinant protein from Escherichia coli using a glutathione affinity culture -- Analysis of purification fractions -- Part IV. Analysis of mRNA levels -- Total RNA purification -- Analysis of gst:egfp mRNA levels by RT-qPCR: Part I -- Analysis of gst:egfp mRNA levels by RT-qPCR: Part II -- Analysis of gst:egfp mRNA levels by semiquantitative RT-PCR, Part I -- Analysis of gst:egfp mRNA levels by semiquantitative RT-PCR, Part II -- Part V. Modulation of gene expression -- Culturing mammalian cells -- Transient transfection of mammalian cells -- RNAi-mediated knockdown of EGFP, Part I -- RNAi-mediated knockdown of EGFP, Part II -- RNAi-mediated knockdown of EGFP, Part III -- CRISPR-mediated knockout of EGFP, Part I -- CRISPR-mediated knockout of EGFP, Part II -- Advanced CRISPR, Part I -- Advanced CRISPR, Part II -- Interim laboratory session -- Advanced CRISPR, Part III -- Advanced CRISPR, Part IV -- Advanced CRISPR, Part V -- Advanced CRISPR, Part VI |
| Summary |
Molecular Biology Techniques: A Classroom Laboratory Manual, Fourth Edition is a must-have collection of methods and procedures on how to create a single, continuous, comprehensive project that teaches students basic molecular techniques. It is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology--or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students will gain hands-on experience on subcloning a gene into an expression vector straight through to the purification of the recombinant protein |
| Notes |
Includes index |
| Bibliography |
Includes bibliographical references and index |
| Notes |
Online resource; title from PDF title page (ScienceDirect, viewed March 14, 2019) |
| Subject |
Molecular biology -- Laboratory manuals
|
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Genetics -- Technique.
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Genetic Techniques
|
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Molecular Biology -- methods
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CRISPR-Cas Systems
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Genetics -- Technique
|
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Molecular biology
|
| Genre/Form |
Laboratory manuals
|
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Laboratory manuals.
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Manuels de laboratoire.
|
| Form |
Electronic book
|
| Author |
Miller, Heather B., author.
|
|
Srougi, Melissa C., author.
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Witherow, D. Scott, author.
|
| ISBN |
9780128157756 |
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0128157755 |
|
