BookSusan Carson, Heather B. Miller, D. Scott Witherow, Melissa C. Srougi.
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.