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cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4108

Category: PCR Protocols: Real-Time PCR Protocols

Protocol describes the real-time PCR using RNA purified from laser-captured tissues Laser Capture Microdissection (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cells. - [Read cDNA Synthesis and Real-Time PCR Using RNA from Laser-Captured Cells Protocol]

Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors - http://www.natureprotocols.com/2006/06/29/cell_staining_for_sorting_of_h.php

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Describes the standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors]

Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA - http://www.natureprotocols.com/2006/06/29/cell_staining_for_sorting_of_h.php

Category: Stem Cell Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Here, we described our standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA]

Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol - http://www.nature.com/nprot/journal/v1/n2/abs/nprot.2006.98.html

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. - [Read Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol]

Collection of Protein Purification Links - http://www.ebioinfogen.com/PROTPURI.htm

Category: Protein Protocols: Protein Purification and Identification

Collection of Protein Purification Links - [Read Collection of Protein Purification Links]

Column Method for Lambda Phage DNA Preparation - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda11.html

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Column Method for Mini-prep of lambda phage DNA purification from lysates. Donis Keller Lab - [Read Column Method for Lambda Phage DNA Preparation]

Column Method for Lambda Phage DNA Preparation Protocol - http://humgen.wustl.edu/hdk_lab_manual/lambda/lambda11.html

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Mini-prep method for lambda phage DNA purification from lysates. - [Read Column Method for Lambda Phage DNA Preparation Protocol]

Comparison Different Protein Quantitation Determination Methods - http://wolfson.huji.ac.il/purification/Protocols/Comparision_of_Methods.htm

Category: Protein Protocols: Protein Quantitation Concentration Protocols

A Comparision of Different Protein Determination Methods and protocols in table format. The Wolfson Centre for Applied Structural Biology The Hebrew University of Jerusalem Dr. Mario Lebendiker. - [Read Comparison Different Protein Quantitation Determination Methods]

Competitive RT-PCR: Preparation of Competitor RNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4110

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

The first step in competitive RT-PCR is the synthesis and purification of the synthetic competitor. This is an RNA molecule designed to be reverse-transcribed and PCR-amplified with the same efficiency as the endogenous transcript of interest. Once the competitor molecule has been prepared, as described in this protocol, competitive PCR can be carried out. - [Read Competitive RT-PCR: Preparation of Competitor RNA Protocol]

Complete Microarray Chip Protocol - http://www.rbhrfcrc.qimr.edu.au/kidney/Pages/Microarray%20protocol.html

Category: DNA Microarray Protocols

Reagents & Equipment, Labelling, Purification, Hybridisation, Washing. KRL Kidney Research Dr.Healt - [Read Complete Microarray Chip Protocol]

Coupling Peptides To Resin For Affinity Purification - Claire Walczak. - http://mitchison.med.harvard.edu/protocols/ab3.html

Category: Peptide Protocols: Antibodies from Peptides Protocols

Coupling Peptides To Resin For Affinity Purification - Claire Walczak. - [Read Coupling Peptides To Resin For Affinity Purification - Claire Walczak.]

Cyanine Dye Purification Protocol Cy - http://cmgm.stanford.edu/pbrown/mguide/hplc.html

Category: DNA Microarray Protocols

Cyanine Dye Purification Protocol using Sep-Pak and HPLC purification protocol for Cy dyes (un)incorporated with dNTP. Brown Lab Howard Hughes Medical Institute - [Read Cyanine Dye Purification Protocol Cy]

Direct Retrieval of DNA Fragments from Pulsed-field Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4083

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for direct retrieval of DNA fragments from pulsed-field gels. A gel slice containing a fragment of DNA resolved by pulsed-field gel electrophoresis is treated with agarase. The released DNA can be used as a substrate for ligation or restriction without further purification. - [Read Direct Retrieval of DNA Fragments from Pulsed-field Gels Protocol]

Direct Selection of cDNAs with Large Genomic DNA Clones Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4077

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

The goal of this method is to identify transcriptionally active genes in cloned segments of genomic DNA. The protocol uses hybridization and affinity purification to recover biotin-labeled cDNAs that bind to a 500-kb segment of human DNA cloned in a BAC vector. However, the method can be easily adapted to other clones of genomic DNAs cloned in high-capacity vectors. - [Read Direct Selection of cDNAs with Large Genomic DNA Clones Protocol]

DNA cloning Procedures - http://hulab.tamu.edu/Protocols/cloning/

Category: DNA Protocols: DNA Cloning

Agarose gel purification, Annealing and extending, oligonucleotides, Ethanol Precipitation, Ligation Miniprep, Oligonucleotide purification, Recovering DNA bands, Restriction digest Gene Clean. The Hu Lab. - [Read DNA cloning Procedures]

DNA Electroelution - http://rothlab.ucdavis.edu/protocols/dna-electroelution.html

Category: DNA Protocols: DNA Extraction Protocols: DNA Extraction Polyacrylamide Gels

DNA Electroelution. This protocol describes the purification of DNA by trapping in a high-salt cushion in a "UEA AnalyticalElectroeluter" (IBI). This machine is no longer manufactured, to our knowledge. However, a smiliar device can be easily made from Plexiglas according to the following diagram, taken from Cornel Mulhardt, Molecular Biology and Genomics (2007) Academic Press, p.52: Schimenti Lab - [Read DNA Electroelution]

DNA Extraction from Agarose Gels Protocol - http://www.methodbook.net/dna/gelextrc.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

DNA Extraction from Agarose Gels Protocol. The page includes cutting out the DNA band from the gel, and describes three methods including 1) Spin-columns (Nucleic acid purification columns), 2) using Dialysis tubing (semi-permeable membrane, Visking tubing), and the 3) Paper strip method.Matt Lewis, Department of Pathology, University of Liverpool. - [Read DNA Extraction from Agarose Gels Protocol]

DNA Fragment Purification from Agarose or Acrylamide - http://axon.med.harvard.edu/~cepko/protocol/mike/D5.html

Category: DNA Protocols: DNA Extraction Protocols: DNA Extraction Polyacrylamide Gels

DNA Fragment Purification from Agarose or Acrylamide. The protocol for fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose or Acrylamide]

DNA Fragment Purification from Agarose or Acrylamide Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/D5.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for DNA fragment purification from agarose or acrylamide. For fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose or Acrylamide Protocol]

DNA Fragment Purification from Agarose Protocol - http://axon.med.harvard.edu/~cepko/protocol/mike/D5.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

DNA Fragment Purification from Agarose Protocol. This protocol is best for fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred. - [Read DNA Fragment Purification from Agarose Protocol]

DNA Purification from DEAE Nitrocellulose Membranes Protocol - http://rothlab.ucdavis.edu/protocols/P01.html

Category: DNA Protocols: DNA Extraction Protocols: Agarose Gel DNA Extraction Protocols

DNA Purification from DEAE Nitrocellulose Membranes Protocol. Jeff Lawrence - [Read DNA Purification from DEAE Nitrocellulose Membranes Protocol]

DNA-Binding Protein Purification with Dynabeads. - http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2671937

Category: DNA Protocols: EMSA DNA-Protein Protocols

Magnetic DNA affinity purification of yeast transcription factor tau--a new purification principle for the ultrarapid isolation of near homogeneous factor. Gabrielsen et al. 1989 - [Read DNA-Binding Protein Purification with Dynabeads.]

Dynal Bead RNA-Binding Protein Purification - http://www.dynalbiotech.com/kunder/dynal/dynalpub401.nsf/$all/0E0753994C78DD9EC1256C5400488E1C

Category: RNA Protocols: RNA-binding Protein Purification

Dynal Bead RNA-Binding Protein Purification. DYNAL. - [Read Dynal Bead RNA-Binding Protein Purification]

E.coli Total RNA Labeling Protocol for High Density Oligonucleotide Array - http://www.genome.wisc.edu/resources/protocols/rnalablelingoligo.htm

Category: E Coli Protocols: E coli Nucleic Acid Protocols

E.coli total RNA labeling protocol for high density oligonucleotide array. Includes: RNA Preparation; Digest RNA and Purification of cDNA; Purify cDNA with Qiaquick PCR purification kit; cDNA Fragmentation and end labeling; Labeling with Terminal Transferase. - [Read E.coli Total RNA Labeling Protocol for High Density Oligonucleotide Array]

EMSA Protocol using 32-p. - http://www.well.ox.ac.uk/flint/EMSA.htm

Category: DNA Protocols: EMSA DNA-Protein Protocols

Primer design, Primer annealing, Primer labelling, Probe purification, In vitro hybridization DNA-protein Binding reaction. Jonathan Flint. - [Read EMSA Protocol using 32-p.]

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Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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