Methods from the Hahn Lab


	Biochemistry Methods Acetone precipitation of protein BioRad Protein Assay Brf1 purification from E. coli Comassie Blue Protein gel stain CNBr protein digest Crosslinking of PICs and IP Fluorescence polarization guide E. coli total protein for SDS PAGE 3x Flag peptide elution Gcn4 purification Gel mobility shift assay (-Mg) Immobilized Template Assay (Pol II) Invitrogen NuPAGE and western transfer Mobile Pilot Plant Ni Sepharose (GE) manual PEAS labeling of Gcn4 Pet_SUMO manual (Invitrogen) Pol II yeast nuclear extract prep (small scale) Pol II in vitro transcription and primer extension assay Pol III in vitro transcription Pol III yeast whole cell extract (small scale) Protein Expression-PET plasmid handbook (Novagen; EMB Biosciences) Silver Stain for SDS PAGE Talon Metal affinity column purification TCA precipitation of proteins TFIIA Purification TFIIIC purification from yeast Yeast Nuclei isolation Yeast protein prep for SDS PAGE and western (rapid) Yeast small scale whole cell extract for IP Western blot Probing and Stripping using dye conjugated antibodies (Odyssey) Western Blot (Li-Cor Biosciences web resources) Western blot probing by Chemiluminescence Genetic Methods Convert yeast A600 to cells/ml EMS mutagenesis of yeast Yeast Plates Yeast Sporulation Yeast spot test assay Yeast transformation with DMSO/LioAc (high efficiency) Yeast transformation (semi rapid) Yeast Methods from the Gottschling lab			Molecular Biology Methods Antibody Production using denatured proteins Chromatin IP assay (Reeves/Tsukiyama) Chromatin IP assay (Fishburn) DNA concentration assay with fluorescent dye DNA sequencing (dye terminator) [FHCRC form] DNA sequencing (rapid Maxam and Gilbert Chemical degradation) DNA Plasmid mini Prep Electrocompetent E. coli preparation Gels native Acrylamide, TBE (35 ml) Gels (DNA sequencing, 100 ml) Gels Acrylamide urea (35 ml gel) in vitro mutagenesis in vitro transcription and translation with PCR DNA Oligonucleotide labeling with ³²P Oligonucleotide purification by PAGE and DEAE PCR with Taq Polymerase (Promega) PCR with expand Polymerase PCR using yeast colonies Primer extension of total Yeast RNA Quantitative measurement of yeast mRNA using RT qPCR qPCR plate and master mix template (power sybr) qPCR 384 well Applied Biosystems SDS triplicate sample template Quickchange mutagenesis with KOD extreme Pol Quickchange site directed Mutagenesis (Pfu Pol) S1 mapping with of yeast RNA with oligonucleotide probe Single stranded dut^- ung^- DNA preparation Yeast DNA preparation (rapid glass bead prep) Yeast genomic DNA Preparation (high quality DNA) Yeast RNA (acid phenol) SM induction and cDNA synthesis Yeast quick plasmid DNA prep Proprietary Methods (password required)

Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N. PO Box 19024 Seattle, WA 98109
©2012 Fred Hutchinson Cancer Research Center, a 501(c)(3) nonprofit organization.
Terms of Use & Privacy Policy.