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2-D Polyacrylamide Gel Electrophoresis Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/2DPage.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

ABI Prism 310 Cycle Sequencing Protocol - http://vosshall.rockefeller.edu/protocols/ABICYCLESEQ.pdf

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for ABI prism 310 cycle sequencing. Includes: Sequencing Reaction; Column Purification; Sample Loading Preparation. - [Read ABI Prism 310 Cycle Sequencing Protocol]

Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb16/16175-2.pdf

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol presents the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies. - [Read Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR]

Analysis of Clones for Exon Trapping and Amplification - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3651

Category: DNA Protocols: Genomic DNA Library Protocols

Amplification and sequencing of the exon-trapped products. - [Read Analysis of Clones for Exon Trapping and Amplification]

Analysis of DNA Methylation using Bisulphite Sequencing Protocol - http://www.methods.info/Methods/DNA_methylation/Bisulphite_sequencing.html

Category: PCR Protocols: Bisulfite Conversion Based PCR Protocols

Protocol for the analysis of DNA methylation using bisulphite sequencing. Method allows precise analysis of methylation in a certain region by converting all nonmethylated cytosines into tymines, while methylated cytosines remain unchanged. This method requires small amount of genomic DNA and therefore seems to be very useful for the analysis of clinical samples, where the material amount is limited.
- [Read Analysis of DNA Methylation using Bisulphite Sequencing Protocol]

Analysis of methylation using bisulphite sequencing - http://www.methods.info/Methods/DNA_methylation/Bisulphite_sequencing.html

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

This method allows precise analysis of methylation in a certain region by converting all nonmethylated cytosines into tymines, while methylated cytosines remain unchanged. Dr. A. Gratchev Methods.info - [Read Analysis of methylation using bisulphite sequencing]

Annealing, Cloning, and Sequencing of shRNA Linkers into pEN_H1 Plasmids Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000230.pdf?pid=PP00000230

Category: RNAi Technology Protocols

Protocol describes the preparation of an shRNA-expressing Gateway entry vector. This process is preceded by the design and synthesis of target gene-specific sense and antisense oligonucleotides which, when annealed, will constitute an shRNA cassette. - [Read Annealing, Cloning, and Sequencing of shRNA Linkers into pEN_H1 Plasmids Protocol]

Autoradiography and Reading of Sequencing Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3588

Category: DNA Protocols: DNA Sequencing Protocols

Protocol describes fixation of a sequencing gel and processingin preparation for autoradiography. - [Read Autoradiography and Reading of Sequencing Gels Protocol]

Bisulfite-PCR for Restriction Analysis and/or Sequencing - http://www.mdanderson.org/departments/methylation/display.cfm?id=8E0AAAAC-FA42-486C-A93F60CD6E1C7E51&method=displayFull&pn=A3F15D82-C9B4-41CD-BA4BE767E50A73D2

Category: Epigenetics and Methylation Protocols

Bisulfite-PCR for Restriction Analysis and/or Sequencing. Protocol written by Jean-Pierre Issa. M.D Anderson Cancer Centre Texas. - [Read Bisulfite-PCR for Restriction Analysis and/or Sequencing]

Bisulfite-PCR for Restriction Analysis and/or Sequencing Protocol - http://www.mdanderson.org/departments/methylation/display.cfm?id=8E0AAAAC-FA42-486C-A93F60CD6E1C7E51&method=displayFull&pn=A3F15D82-C9B4-41CD-BA4BE767E50A73D2

Category: PCR Protocols: Bisulfite Conversion Based PCR Protocols

Protocol for bisulfite-PCR for restriction analysis and/or sequencing. Bisulfite-PCR followed by restriction is a rapid and semi-quantitative method of analyzing DNA methylation. The PCR products are also suitable for either direct sequencing or cloning and sequencing. The most important step here is primer selection. - [Read Bisulfite-PCR for Restriction Analysis and/or Sequencing Protocol]

Carboxypeptidase-Y C-terminal sequencing - http://prowl.rockefeller.edu/recipes/cproteas/carboxyy.htm

Category: Mass Spectrometry Protocols

Carboxypeptidase-Y C-terminal sequencing and sequencing ladder protocol method. PROWL - [Read Carboxypeptidase-Y C-terminal sequencing]

Chemical Sequencing Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3804

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for chemical sequencing. - [Read Chemical Sequencing Protocol]

Construction of the Sequencing Library Protocol - http://sequence-www.stanford.edu/protocols/library.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol first describes the vector preparation and, then, describes the insert preparation. Vital to have an excellent vector in order to produce a sequencing library. Protocol employs the male-specific coliphage M13 as the sequencing vector. M13 is a filamentous phage with a single-stranded, circular genome. M13 is widely used as a vector because many versions are available commercially and because M13 has certain advantages. - [Read Construction of the Sequencing Library Protocol]

Cutting Bands and Reamplification for Sequencing Protocol - http://genome-lab.ucdavis.edu/Protocols/Cutting%20Bands%20and%20Reamplification%20for%20Sequencing.htm

Category: Genetics and Genomics: Cancer Genetics Protocols

Protocol for cutting bands and reamplification for sequencing. - [Read Cutting Bands and Reamplification for Sequencing Protocol]

Cycle Sequencing for PCR Product Protocol - http://genoseq.ucla.edu/action/view/PCR_product_Cycle_Sequencing

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for cycle sequencing for PCR product. - [Read Cycle Sequencing for PCR Product Protocol]

Dideoxy-mediated Sequencing of DNA Using Taq DNA Polymerase Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3803

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Because sequencing reactions catalyzed by thermostable DNA polymerasessuch as Taq are carried out at elevated temperatures, problemscaused by mismatched annealing of primers or templates richin secondary structure are greatly alleviated. - [Read Dideoxy-mediated Sequencing of DNA Using Taq DNA Polymerase Protocol]

Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3801

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol describes DNA-sequencing of single-stranded DNAtemplates in reactions catalyzed by Sequenase. - [Read Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol]

Dideoxy-mediated Sequencing Reactions Using PCR and End-labeled Primers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3791

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

In this protocol asymmetric PCR is used to generate single-strandedDNA templates for dideoxy-sequencing - [Read Dideoxy-mediated Sequencing Reactions Using PCR and End-labeled Primers Protocol]

Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3802

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol for dideoxy-mediated sequencing reactions using the Klenow fragment of E. coli DNA polymerase I and single-stranded DNA templates. - [Read Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase]

Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase I - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3802

Category: E Coli Protocols: E coli Nucleic Acid Protocols

Protocol for dideoxy-mediated sequencing reactions using the Klenow Fragment of E. coli DNA polymerase I and single-stranded DNA templates. - [Read Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase I]

Direct Sequencing Using MultiPlex PCR-Based Method - http://www.genome.ou.edu/MultiplexPCRbasedSeq.html

Category: DNA Protocols: DNA Sequencing Protocols

Direct sequencing from amplified bacterial and large insert cloned human or mouse genomic DNA via an improved MultiPlex PCR-based method. Includes: Preparing primers for MP-PCR; Amplification; PCR Product Clean-up; Sequencing. - [Read Direct Sequencing Using MultiPlex PCR-Based Method]

DNA Sequencing Methods - http://www.genome.ou.edu/protocol_book/protocol_partIV.html#IV.C

Category: DNA Protocols: DNA Sequencing Protocols

Methods for DNA sequencing. Includes: Bst-catalyzed radiolabeled DNA sequencing; Radiolabeled sequencing gel preparation, loading, and electrophoresis; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye terminator reactions; Sequenase[TM] catalyzed sequencing with dye-labeled terminators; etc. - [Read DNA Sequencing Methods]

Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates - http://www.genome.ou.edu/big_dyes_cos_bac_pac_fos.html

Category: BACs Bacterial Artificial Chromosome Protocols

Sequencing conditions tested for the ABI Big-Dye terminators (PE-ABI #4303150 for the 1000 reaction kit - Description: TF,KIT BTD RR-1000) with various templates. Important to quantitate all templates by agarose gel electrophoresis vs size and concentration standards and do a few tests with different template concentrations to determine the optimal conditions for your reactions. Several conditions are given. - [Read Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates]

Dye Terminator Cleanup for Automated DNA Sequencing Reactions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4101

Category: DNA Protocols: DNA Sequencing Protocols

Protocol describes a simple procedure for purifying sequencing reactions using the MagneSil particles from Promega. - [Read Dye Terminator Cleanup for Automated DNA Sequencing Reactions Protocol]

Embryo Lysates & Immunoprecipitation Protocol - http://spot.colorado.edu/~klym/Methods/Precipitation.htm

Category: Xenopus Protocols: Xenopus Embryo Manipulation Protocols

Protocol for embryo lysates and immunoprecipitation: protein A agarose & protein A magnetic beads. Includes: Embryo lysates; Immunoprecipitation with protein A agarose; Immunoprecipitation with protein A magnetic beads for silver stain/sequencing analysis. - [Read Embryo Lysates & Immunoprecipitation Protocol]

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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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