McManusLab dissecting the dark matter of our genome
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McManus Lab Protocols

General Cell Biology
Thawing ES cells cpark
Staining for beta-galactosidase activity cpark
Rubidium Chloride Competent Cell Protocol hueyjiin
Picking ES cell colonies and transferring them to 96-well plates cpark
Passaging/Splitting 96-well Master Plate into Three Plates cpark
Lac Z staining mouse embryos cpark
Growing/splitting E14 ES cells cpark
Freezing ES cells cpark
Freezing 96-well master plates cpark
Electroporation of targeting construct cpark
DNA Isolation from ES Cells -96 well plate cpark
Culturing Immortalized Dicer Floxed MEFs hunter
24-well Plate Spin Infection tchan

microRNA
Small RNA Northern Blotting Trinna22
miRNA microarrays gloeb
Library cloning protocol - mir30 context wpatena

General Molecular Biology
Site-Directed Mutagenesis (Stratagene protocol) jkolibachuk
Preparing 96-well Genotyping Plate for genomic DNA Extraction cpark
P32 Probe Preparation ychoi
MicroRNA and siRNA Cloning Protocol ychoi
Metabolic Labeling Using 32P ychoi
Making Long dsRNA from In Vitro Transcriptions ychoi
Cloning via Restriction Digest Carol R
cip Carol R
Chemical Synthesis of ImpA (adenosine 5'-phosphorimidazolide) ychoi
A PCR-based strategy for cloning short hairpin sequences: “PCR shagging" mmcmanus
8-cell Embryo Collection mmcmanus

RNAi
siRNA transfection with HiPerfect (MIN6 cells) gregku
PCR template method for dsRNA Synthesis zli
Lymphocyte siRNA electroporation transfection mmcmanus
Lipid siRNA transfection mmcmanus
Library cloning protocol - shRNA context wpatena
Creating RNAi stem loops for LentiLox 3.7 mmcmanus

Virus
Lentivirus harvest and concentration xwang
Lentiviral Vectors Perivitelline Injection zli
Lentiviral Transfection Protocol for a 10cm dish xwang
Lentiviral Transfection xwang
Lentiviral Harvest and Concentration xwang
LentiLox Vector Design mmcmanus
CLONING SMALL HAIRPINS INTO LENTIVIRAL VECTORS rlebbink