Biochemistry

Acetone precipitation of protein
BioRad Protein Assay
Comassie Blue Protein gel stain
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
LiCOR Western Blot Normalization
LiCor Western blot quantitation and troubleshooting
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)
Western blot quantitation and troubleshooting (LiCor)

Protein Purification Handbooks (GE):

Affinity Chromatography-Antibodies
Affinity Chromatography-tagged proteins
Affinity Chromatography-biomolecules
Ion Exchange Chromatography
Size Exclusion Chromatography
Design of Protein Purification
SUMO-tag protein purification
TCA precipitation of proteins
Yeast protein prep for SDS PAGE (NaOH; rapid)
Western blot Probing using dye conjugated antibodies (Odyssey)

Molecular Biology

Antibody Production using denatured proteins
Chec-Seq to map genome-wide protein-DNA binding in yeast
DNA purification with AMPure beads (manual)
DNA size selection with AMPure beads (manual)
DNA sequencing (dye terminator) [Hutchbase]
DNA sequencing (rapid Maxam and Gilbert Chemical degradation)
Electrocompetent E. coli preparation
Gels native Acrylamide, TBE (35 ml gel)
Gels, DNA sequencing (100 ml gel)
Gels Acrylamide urea (35 ml gel)
in vitro mutagenesis with dut ung DNA
in vitro transcription and translation with PCR DNA
KOD extreme Polymerase
Oligonucleotide labeling with ³²P ATP
Oligonucleotide purification by PAGE and DEAE
PCR with expand Polymerase
PCR using yeast colonies (KOD extreme)
PicoGreen dsDNA assay
Primer extension of total Yeast RNA
Protocols.io
Qbit Protein assay (full instructions) or Quick Reference
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 for multiple mutations (Megaprimed QCM)
Quickchange site directed Mutagenesis (Pfu Pol)
RNA: 4Thio Uracil RNA labeling and purification
S1 mapping with of yeast RNA with oligonucleotide probe
Single stranded dut- ung- DNA preparation
Transfer PCR (PCR cloning) [Ariel]
Yeast DNA preparation (rapid glass bead prep)
Yeast RNA (acid phenol) SM induction and cDNA synthesis

Yeast Genetic Methods

BPA yeast Media
Convert yeast A600 to cells/ml
Coulter counter Z2 instructions
Coulter counter preference file (Shawna Miles)
EMS mutagenesis of yeast
Yeast Plates
Yeast Sporulation
Yeast spot test assay
Yeast transformation with DMSO/LioAc (high efficiency)
Proprietary Methods (password required)