Lab Protocols

 

The below are somewhat random, and are added upon request. Let us know if you'd like to see a specific protocol posted.

General Molecular Biology

Preparation of high titer electrocompetent cells

Actin Cre genotyping

LacZ Genotyping

8-cell Embryo Collection

Rubidium Chloride Competent Cell Protocol

P32 Probe Preparation

MicroRNA and siRNA Cloning Protocol

Making Long dsRNA from In Vitro Transcriptions

A PCR-based strategy for cloning short hairpin sequences: “PCR shagging"

CIP

Cloning via Restriction Digest

Site-Directed Mutagenesis (Stratagene protocol)

Chemical Synthesis of ImpA (adenosine 5'-phosphorimidazolide)

Metabolic Labeling Using 32P

Preparing 96-well Genotyping Plate for genomic DNA Extraction

General Cell Biology

NIH3T3 transfection with Jetprime Polyplus

Culturing Immortalized Dicer Floxed MEFs

Staining for beta-galactosidase activity

Lac Z staining mouse embryos

Freezing ES cells

Thawing ES cells

Growing/splitting E14 ES cells

Electroporation of targeting construct

Picking ES cell colonies and transferring them to 96-well plates

Passaging/Splitting 96-well Master Plate into Three Plates

Freezing 96-well master plates

DNA Isolation from ES Cells -96 well plate

24-well Plate Spin Infection

Virus

CLONING SMALL HAIRPINS INTO LENTIVIRAL VECTORS

Lentiviral Transfection Protocol for a 10cm dish

Lentiviral Harvest and Concentration

LentiLox Vector Design

Lentiviral Vectors Perivitelline Injection

Lentivirus harvest and concentration

Lentiviral Transfection

RNAi

Lipid siRNA transfection

PCR template method for dsRNA Synthesis

Lymphocyte siRNA electroporation transfection

Library cloning protocol - shRNA context

siRNA transfection with HiPerfect (MIN6 cells)

microRNA

miRNA microarrays

Small RNA Northern Blotting

Library cloning protocol - mir30 context