Web
sites
Biotechniques Molecular Biology Forum
Molecular biology
PCR
- basic method
Reverse
transcription (Qiagen Omniscript)
PCR,
QPCR and sequencing primers
(Old primer dilution method)
Real-time
RT-PCR with SYBR green
Purification of PCR
product by ExoSAPit
Purification of short (<200
bp) PCR products - Linear acrylamide
Agarose
gel electrophoresis
Agar
plate preparation
Trizol protocol for RNA extraction
Genomic
DNA prep by DNAzol
DNAzol
protocol for DNA extraction
Blood
and tissue DNA by Qiagen column
DNA
concentration by Yo-Pro fluorescence
RNA
and DNA concentration by UV absorbance
Firefly
luciferase reagent preparation (homebrew)
Firefly/renilla Promega DualGlo assay protocol in 96 well
DNA extraction from whole blood by salt ppt
Plasmid
mutagenesis with DpnI (Stratagene Quikchange)
DNA
extraction from FTA cards
Western blotting - UGT1A6
General Western blotting protocol
UGT1A6 exon 1 PCR
UGT1A3'-UTR PCR
UGT real-time PCR primer sets
Cell culture
Selection
of cells with Geneticin (G418)
Transient
transfection and stable cell lines - HEK293
Freezing and
thawing eukaryotic cells - protocol
Lipofectamine 2000 general transfection protocol
Lipofectamine
2000 cotransfection plasmid RNAi
Beta
gal assay - Promega
Some good cell culture practices
Caco2 protocol
LS180 protocol
Microsomes, activities,
TLC, and HPLC
Liver
microsomes preparation
Protein
concentration assay by BCA method (Pierce)
Protein (BCA) assay on microtiter plate
UDPGA
and NADPH cofactor solutions
UGT activity
screen by C14UDPGA-TLC
Development
of UGT activity assay by HPLC and preliminary phenotyping
Glucuronidase confirmation
of glucuronide metabolite
Deconjugation of
drug metabolites in urine
Measurement
of soy isoflavones in pet food
HPLC-mass spec buffer preparation
Triazolam in plasma by LCMS
Other methods
Calibration
of Ohaus scale
Maintenance of ABI 7300 Real-time PCR machine
QuickCalcs online calculator
Simple Hardy-Weinberg
Calculator - Court Lab
Comparative and Molecular Pharmacogenomics Laboratory
Department of Veterinary Clinical Sciences,
Washington State University College of Veterinary Medicine
100 Grimes Way, Pullman, WA 99163,
USA.
Phone: 509-335-0817; Fax: 509-335-0880
Comments or problems please send email to: michael.court@vetmed.wsu.edu
Last update: 10 October, 2012