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Nuclear Extract Preparation Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/N1.html

Category: DNA Protein Interactions Protocols

Protocol for nuclear extract preparation. - [Read Nuclear Extract Preparation Protocol]

Oligonucleotide Purification Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/O1.html

Category: Nucleic Acid Purification Protocols

Protocol for oligonucleotide purification. - [Read Oligonucleotide Purification Protocol]

PCR Genotyping from Tail DNA Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/P4.html

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Protocol for PCR genotyping from tail DNA. This protocol works well for a variety of genes and primer pairs including Tg and KO alleles. Oligonucleotide melting temperatures between 60° and 65° seem to work well. - [Read PCR Genotyping from Tail DNA Protocol]

PCR Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/P3.html

Category: PCR Protocols: Standard PCR Protocol

Protocol for PCR. - [Read PCR Protocol]

Plasmid Sequencing Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S1.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for plasmid sequencing. - [Read Plasmid Sequencing Protocol]

Preparation Of Peptide-KLH Conjugates For Immunization - http://mitchison.med.harvard.edu/protocols/ab1.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Preparation Of Peptide-KLH Conjugates For Immunization. Coupling of Peptide to KLH. And generation of Antibodies using peptide-KLH conjugate - Claire Walczak. - [Read Preparation Of Peptide-KLH Conjugates For Immunization]

Preparation Of Peptide-KLH Conjugates For Immunization - http://mitchison.med.harvard.edu/protocols/ab1.html

Category: Immunology

Checking Peptide Thiol Groups, Coupling of Peptide to KLH. Claire Walczak - [Read Preparation Of Peptide-KLH Conjugates For Immunization]

Preparation of Segmented and Polarity Marked Microtubules Protocol - http://mitchison.med.harvard.edu/protocols/polarity.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Segmented and polarity-marked microtubules are very useful for many different types of in vitro assays. Segmented microtubules are microtubules with a bright seed and dim elongated segments on both ends. Polarity marked microtubules are microtubules with a bright seed and a dim elongated segment only on one end -- the plus end. - [Read Preparation of Segmented and Polarity Marked Microtubules Protocol]

Printing proteins as microarrays for high-throughput function determination. - http://cgr.harvard.edu/macbeath/protocols/protein_microarrays.html

Category: Protein Microarray Protocols

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

Protein Microarray Information - http://cgr.harvard.edu/macbeath/research/protein_ma.html

Category: Protein Microarray Protocols

Protein Microarray Information. MacBeath Lab - [Read Protein Microarray Information]

Protocol for Real-Time RT-PCR Using SYBR Green - http://pga.mgh.harvard.edu/primerbank/PCR_protocol.html

Category: Real Time PCR

A detailed experimental procedure for real-time RT-PCR using SYBR Green I as mentioned in Wang et al 2003. - [Read Protocol for Real-Time RT-PCR Using SYBR Green]

Protocol for Stable Lines - http://genetics.med.harvard.edu/~cepko/protocol/mike/S4.html

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol is optimized for erythroid cells, other cell types may require modifications. - [Read Protocol for Stable Lines]

Purification Of Anti Peptide Antibody - Claire Walczak - http://mitchison.med.harvard.edu/protocols/ab4.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Purification Of Anti Peptide Antibody - Claire Walczak - [Read Purification Of Anti Peptide Antibody - Claire Walczak]

Quick DNA Plasmid Prep Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D4.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Protocol for quick DNA plasmid prep. Protocol gives very clean plasmid preps for restriction digests and cloning. However, due to the alkaline lysis step, the DNA is often nicked and may not give exceptional sequence data. - [Read Quick DNA Plasmid Prep Protocol]

Quick DNA Plasmid Prep. Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D2.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Protocol for quick DNA plasmid prep. This is a very fast mini-prep protocol which is suitable for sequence analysis and restriction digests. - [Read Quick DNA Plasmid Prep. Protocol]

Random Prime Labeling of DNA Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R1.html

Category: Nucleic Acid Modification Protocols: Random Primer Labeling Protocols

Protocol for random prime labeling of DNA. - [Read Random Prime Labeling of DNA Protocol]

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

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

Long PCR Buffer, Cycle times and temperatures, Picking Primers, Hot Start Polymerases for Long PCR. Harvard-Lipper Center for Computational Genetics. Modified from Cheng et al. - [Read Reagents and Guidelines for Long PCR]

Recycling Tubulin Protocol - http://mitchison.med.harvard.edu/protocols/recycle.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Recycle tubulin fractions stored at -80¡C after the PC column and store the recycled tubulin in small aliquots for day-to-day use. Generally store recycled tubulin in Injection Buffer (IB) without free GTP. This is done because depolymerization appears to be much better in IB, IB is ideal for microinjections/adding tubulin to extracts, and the absence of free GTP makes polymerization with GMPCPP, a very useful GTP analog that has ~5-10X lower affinity than GTP for tubulin. - [Read Recycling Tubulin Protocol]

Removal of Small Fragments of Nucleic Acid from Preparations of Plasmid DNA by Centrifugation - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3916

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Contamination of plasmid DNA by fragments of DNA and RNA is reduced to an acceptable level by centrifugation through 1 M sodium chloride. This method was devised by Brian Seed when he was a graduate student at Harvard University. - [Read Removal of Small Fragments of Nucleic Acid from Preparations of Plasmid DNA by Centrifugation]

RNA Preparation from Cultured Cells or Tissue Samples Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R2.html

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols: RNA Isolation fromTissues or Cultured Cells Protocols

Protocol for RNA preparation from cultured cells or tissue samples. This protocol has been used to isolate RNA from relatively small tissue samples. The RNA is clean enough for Rnase protection, cDNA synthesis, and RT-PCR analysis. - [Read RNA Preparation from Cultured Cells or Tissue Samples Protocol]

RNA Whole Mount In Situ Hybridization Protocol - http://genetics.med.harvard.edu/~cepko/protocol/ctlab/ish.ct.htm

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for RNA whole mount in situ hybridization. Includes: Embryo preparation; Prehybridization and Hybridization; Post-hybridization washes, blocking, and antibody incubation; Post-antibody washes; Color development. - [Read RNA Whole Mount In Situ Hybridization Protocol]

Rnase Protection Assay Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R3.html

Category: Molecular Biology Protocols

Protocol for RNAse protection assay. - [Read Rnase Protection Assay Protocol]

RT-PCR Analysis Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R4.html

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Protocol for RT-PCR analysis. - [Read RT-PCR Analysis Protocol]

Silinization of Slides Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S6.html

Category: Immunohistochemistry Protocols: Coated Treated Slides Protocols

Protocol for silinization of slides. - [Read Silinization of Slides Protocol]

Single Strand DNA Prep for Sequencing Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S2.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for single strand DNA prep for sequencing. Protocol works well from a 5 ml starting culture or a 1 ml starting culture. - [Read Single Strand DNA Prep for Sequencing Protocol]

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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