KIELKOPF LAB PROTOCOLS


Files are in PDF format and are password protected.


General

Perkin Elmer Lambda25 UV/ VIS Spectrophotometer

Turning off UV/ VIS Spectrophotometer Lamps

Washing Glassware

Using Autoclave

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Subcloning protocols

Making LB for Plates and Liquid Media

Antibiotics Guide

Single site mutagenesis

PCR Primer Design for Subcloning

PCR

Restriction Enzyme Digest of DNA Insert and Vector

Gel Purifying Your PCR Product

Photographing DNA gels

Ligation

Bacterial Transformation

Colony PCR

Overnight Cultures of successful transformants

DNA prep for sequencing

DNA sequencing at UR

Freezer stocks of transformed bacteria

Making Competent Cells

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Protein Purification and Biochemistry Protocols

Care for Akta Purifiers

E. coli Large Scale Induction of Protein Expression

Lysis of E. coli by Sonication

Protein Purification Buffers

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

Glutathione Resin / Agarose Preparation

Expression Test

GST pull-down

GST competition pull-down

Precision Protease Purification and Use


U2AF65 DLink Purification

SF1 Purification

SF1 Mystery Domain Purification

Full length histag-U2AF65 Purification

Full length GST-ATPase Purification

TIA-1 RRM123 Purification

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Nucleic Acid Protocols

DNA Desalting

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Thermodynamic and Structural Protocols

Isothermal titration calorimetry (ITC)

Circular Dichroism Protocol

Fluoromax-3 polarizer alignment

Equations for fitting fluorescence titration assay

Dynamic light scattering (DLS)

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

Surface Plasmon Resonance using Reichert SPR

R-axis IV login information

Crystal Photos

Effect of interdomain linker on Kd

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