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URMC / Labs / Kielkopf Lab / Lab Protocols


Lab Protocols


Nanodrop UV/VIS Spectrophotometer

Lab Key Code

Washing Glassware - Manual

Washing Glassware - Dishwasher

Using Autoclave

Protein Purification and Biochemistry Protocols

Care for Akta Purifiers

Akta Fraction Collector Racks

Remote Connection to Akta Pures

E. coli Large Scale Induction of Protein Expression

Lysis of E. coli by Sonication

Lysis of E. coli by Avestin press

Protein Purification Stock Buffers

Bacterial Cell Resuspension Buffers

Protein Purification Running Buffers

Purify a Protein with a GST Tag

Purify a Protein with a Histidine Tag

Purify a Protein with Acidic pI

Purify a Protein with Basic pI

Purify a Protein with Thrombin Cleavage Site

Selenomethionine Protein Purification

A280 Protein Concentration Determination

Dialysis Tubing

Laemmli Method for SDS-PAGE Gels and Buffers

Tricine Gels

Prepare Sigma glutathione agarose

Expression Test

GST Pull-Down

GST Competition Pull-Down

PreScission Protease Purification and Use

TEV Protease Purification and Use

U2AF65 DLink Purification

SF1 Purification

U2AF65 UHM Purification

Nucleic Acid Protocols

DNA Desalting 2g SepPak

DNA Desalting 200mg SepPak

Nitrocellulose Filter Binding

Electrophoretic Mobility Shift Assays

Adjusting PhosphoImager Image Quality

SYBR Gold Staining of Nucleic Acids

Thermodynamic Protocols

BIAcore T100 for kinetic or affinity measurements

Isothermal Titration Calorimetry using Kielkopf VP-ITC

Isothermal Titration Calorimetry using Wedekind PEAQ-ITC

Circular Dichroism Protocol: Jasco in SBBF

Circular Dichroism Protocol: Prior Aviv

Fluoromax-3 Polarizer Alignment

Fluorescence Anisotropy RNA Binding Assay

Using Fluoromax-4 Fluorimeter

Equations for Fitting Fluorescence Titration Assay

Electrophoretic Mobility Shift Assay

Amino Acid Analysis to Confirm Concentration for ITC

Effect of Interdomain Linker on Kd

Structural Biology Protocols

Vitrobot for cryoEM Grid Preparation

Dynamic Light Scattering (DLS)

Small Angle X-ray Data Collection at ALS (with Greg Hura's help)

Scaling Pr functions calculated from SAXS data

Running MONSA on SAXS data

Bruker CCD login information

Crystal Photos

Crystal Flash-Cooling Tips

Remote Data Collection at SSRL

Prepare SeMet Protein

Sazaki Table of f' and f"

Molecular Biology Protocols

Making LB for Plates and Liquid Media

Antibiotics Guide

Bacterial Transformation

Single site mutagenesis

PCR Primer Design for Subcloning


Restriction Enzyme Digest of DNA Insert and Vector

Gel Purifying Your PCR Product

Analyzing Plasmid DNA by Gel Electrophoresis

Photographing DNA Gels


Colony PCR

Overnight Cultures of Successful Transformants

DNA prep for sequencing

DNA sequencing at UR

Freezer stocks of transformed bacteria

Making Competent Cells

Tissue Culture and Related Protocols

Endotoxin-free Maxipreps

Tissue Culture Guide

Quantitative Real-Time Reverse Transcription-PCR

Immunoblot Protocol

Mary Pulvino’s coIP videos (courtesy of Justin Galardi)


In Vitro Splicing Assay

Quantifying RT-PCR images