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Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol - http://genetics.med.harvard.edu/~cepko/electroporation/PNAS-final.pdf

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol for electroporation and RNAi in the rodent retina in vivo and in vitro. - [Read Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol]

Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol - http://genetics.med.harvard.edu/~cepko/electroporation/PNAS-final.pdf

Category: RNAi Technology Protocols

Protocol for electroporation and RNAi in the rodent retina in vivo and in vitro. - [Read Electroporation and RNAi in the Rodent Retina In Vivo and In Vitro Protocol]

EMSA using ds Oligonucleotides - http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html

Category: DNA Protocols: EMSA DNA-Protein Protocols

EMSA probe creation using ds Oligonucleotides by annealing two complementary oligos, and Klenow for probe creation. - [Read EMSA using ds Oligonucleotides]

EMSA using ds Oligonucleotides Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/E1.html

Category: DNA Protein Interactions Protocols: Electrophoretic Mobility Shift Assays EMSA Protocols

Protocol for EMSA using ds oligonucleotides. - [Read EMSA using ds Oligonucleotides Protocol]

Experimental Procedures for Protein Microarrays - http://web.archive.org/web/20050313184601/http://cgr.harvard.edu/macbeath/protocols/proteinmicroarrays.html

Category: Protein Microarray Protocols

Experimental Procedures for Protein Microarrays. MacBeath G & Schreiber SL (2000). Printing proteins as microarrays for high-throughput function determination. Science 289: 1760-1763. - [Read Experimental Procedures for Protein Microarrays]

Expression and Purification of His Tagged Proteins in E. coli Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/H1.html

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression and purification of His tagged proteins in E. coli. - [Read Expression and Purification of His Tagged Proteins in E. coli Protocol]

Fill-in Labeling of DNA Fragments Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F1.html

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Protocol for fill-in labeling of DNA fragments. - [Read Fill-in Labeling of DNA Fragments Protocol]

Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Protocol - http://mitchison.med.harvard.edu/protocols/flowcell.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Protocol for flow cell assays with microtubules. Includes: Solutions & Supplies; VE-DIC Assays; Motility; Dynamics; Fluorescence Assays; Oxygen Scavenging; Assays. - [Read Flow Cell Assays with Microtubules: Motility/Dynamics in Fluorescence and VE-DIC Protocol]

Flow Cytometry Analysis Protocol - http://cbr.med.harvard.edu/labs/springer/protocols/qing_facs.html

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Flow Cytometry Analysis Protocol. Springer Lab - Harvard. Background on flow cytometry technique: Flow cytometry is a method which uses instrumentation scanning single cells flowing past excitation sources in a liquid medium.
- [Read Flow Cytometry Analysis Protocol]

Frozen Competent E. coli Preparation and Transformation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F3.html

Category: E Coli Protocols: E coli Transformation Protocols

Protocol for frozen competent E. coli preparation and transformation. - [Read Frozen Competent E. coli Preparation and Transformation Protocol]

Frozen Competent E. coli Preparation and Transformation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F3.html

Category: Competent Cells Protocols

This is a very easy protocol which yields transformation efficiencies appropriate for routine cloning and transformation. - [Read Frozen Competent E. coli Preparation and Transformation Protocol]

High Efficiency FCC Preparation and Tx Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/F4.html

Category: Competent Cells Protocols

This is an easy and straightforward protocol that gives efficiencies of 106 to 107 cfu/mg of plasmid DNA. A growth curve is required for each strain that is prepared.
- [Read High Efficiency FCC Preparation and Tx Protocol]

Histochemical Staining Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/H2.html

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Protocol for histochemical staining. This protocol has been optimized for b-galactosidase and human placental alkaline phosphatase staining in retinal tissue and cultured cells. - [Read Histochemical Staining Protocol]

HSS Depletion Conditions for XKCM1 Protocol - http://mitchison.med.harvard.edu/protocols/ext2.html

Category: Xenopus Protocols: Xenopus Egg Extracts Protocols

Protocol for HSS depletion conditions for XKCM1. - [Read HSS Depletion Conditions for XKCM1 Protocol]

Lambda DNA Preparation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D3.html

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes a plate method that gives very good yield for cloning. Includes: T-TYN Media + Mg+2; T-TYN Plates; T-TYN Top Agarose. - [Read Lambda DNA Preparation Protocol]

Lambda DNA Preparation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D3.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for lambda DNA preparation. This is a plate method that gives very good yield for cloning. - [Read Lambda DNA Preparation Protocol]

Lambda gt22a Screening Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/L1.html

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol for lambda gt22a screening. Includes: T-TYN Media + Mg+2; T-TYN Plates; T-TYN Top Agarose; Phage Dilution Buffer (PDB); 10X Kinase Buffer (-DTT); Calf Thymus DNA. - [Read Lambda gt22a Screening Protocol]

Lineage Analysis Using Retroviral Vectors - http://genetics.med.harvard.edu/~cepko/lineage/

Category: Developmental Biology

Describe the use of replication-incompetent retroviral vectors for the analysis of lineal relationships in developing vertebrate tissues. An overview of the relevant aspects of the retroviral life cycle, and the strategies and current methods in use at their laboratory are described. - [Read Lineage Analysis Using Retroviral Vectors]

Lineage Analysis Using Retroviral Vectors Protocol - http://genetics.med.harvard.edu/~cepko/lineage/

Category: Cloning Protocols: Vector Protocols

Protocol describes the use of replication-incompetent retroviral vectors for the analysis of lineal relationships in developing vertebrate tissues. - [Read Lineage Analysis Using Retroviral Vectors Protocol]

Long PCR Reagents and Guidelines - http://arep.med.harvard.edu/labgc/estep/longPCR_protocol.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols: Long PCR Protocols

General Guidelines for Long PCR Conditions and Enzyme Mixtures. Pete Estep. Harvard. - [Read Long PCR Reagents and Guidelines]

Lysate Microarrays - Information - http://cgr.harvard.edu/macbeath/research/lysate_ma.html

Category: Protein Microarray Protocols

Lysate Microarrays. High throughput methods that detect abundance and post-translational modification. MacBeath Lab - [Read Lysate Microarrays - Information]

Maxam Gilbert Sequencing for DnaseI Footprinting Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/M2.html

Category: DNA Protein Interactions Protocols: Dnase Footprinting Assay Protocols

Quick protocol to give a representative G+A ladder for DnaseI footprint experiments. - [Read Maxam Gilbert Sequencing for DnaseI Footprinting Protocol]

Methylation Interference Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/M1.html

Category: DNA Protein Interactions Protocols

Protocol for methylation interference. - [Read Methylation Interference Protocol]

Negative Stain Electron Microscopy of Microtubules Protocol - http://mitchison.med.harvard.edu/protocols/EM.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Negative staining is a rapid, qualitative method for analyzing microtubule structure at the EM level. Because negative staining involves deposition of heavy atom stains, structural artifacts such as flattening of the cylindrical microtubule and opening up of microtubules into flat sheets are common. Negative staining is very useful because of its ease, rapidity and lack of requirement for specialized equipment other than that found in a regular EM facility. - [Read Negative Stain Electron Microscopy of Microtubules Protocol]

Nuclear Extract Preparation for EMSA (PDF) - http://research.dfci.harvard.edu/shivdasani/protocols/Nuclear_extract_prep.pdf

Category: DNA Protocols: EMSA DNA-Protein Protocols

Nuclear Extract Preparation for EMSA after Wadman et al., EMBO J. 1997. Buffer recipies included. - [Read Nuclear Extract Preparation for EMSA (PDF)]

Articles (1-2 of 2)

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

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