Protocol Matches: 282

• TA Cloning (Hubbard Center for Genome Studies)
  Includes procedures for PCR, ligation, transformation and colony screening
  http://hcgs.unh.edu/protocol/basic/Cltaclone.html
  Added: Fri Feb 06 2009, Hits: 3166, Reviews: 0  Write review   Cached
• Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) (Reddy research laboratory, Neurological Sciences Institute, Oregon Health & Science University)
  PCR Method for mutation detection.
  http://www.ohsu.edu/nsi/faculty/reddyh/lab/protssc...
  Added: Mon Sep 23 2002, Hits: 1304, Reviews: 0  Write review   Cached
• RT-PCR Protocol (Julie B. Wolf, University of Maryland, Baltimore County)
  First strand synthesis and subsequent PCR amplification. Based on LTI protocol
  http://www.research.umbc.edu/~jwolf/m12.htm
  Added: Tue May 14 2002, Hits: 8051, Reviews: 0  Write review   Cached
• C. Elegans Gene Knockout (Vancouver Gene Knockout Laboratory, at the University of British Columbia)
  The procedure is to mutagenize a large population of worms with trimethylpsoralen and UV irradiation, set up 1152 subpopulations, screen DNA made from this library for deletions in specific genes by nested PCR, and then to recover single worms carrying the deletions through a sib-selection process.
  http://ko.cigenomics.bc.ca/protocols.html
  Added: Tue May 14 2002, Hits: 439, Reviews: 0  Write review   Cached
• Rapid Extraction of High Quality DNA from Whole Blood Stored at 4ºC for Long Period (Vahid Iranpur-mobarakeh, Department of Animal Science, Faculty of Agriculture, Shahrekord University, Shahrekord, Iran)
  None of the procedures yielded DNA of suitable purity for SSR and other PCR assays for DNA that was stored at 4ºC on prolonged period. We established an improved procedure for rapid isolation of DNA from sheep’s blood and other species stored at 4ºC for up to one year or more and suitable for SSR analysis and other PCR-based applications
  Added: Sun May 02 2010, Reviews: 0  Write review  
• Extraction of genomic DNA from whole blood (Laura-Lee Boodram, Department of Life Sciences, The University of the West Indies)
  The protocol is simple and fairly rapid. It does not require the use of organic solvents but rather utilizes salt extraction to precipitate contaminating proteins. High quality DNA is obtained suitable for immediate PCR applications. One can obtain approximately 100-200 ug of DNA from 4-8 mL of fresh or frozen whole blood.
  Added: Mon May 17 2004, Reviews: 0  Write review  
• DNA Methylation Analysis by Bisulfite Sequencing (BS) (Epigenome Network of Excellence)
  Bisulfite modification converts unmethylated cytosine to uracil, while methylated cytosine does not react. After denaturation and bisulfite modification, double-stranded DNA is obtained by primer extension and the fragment of interest is amplified by PCR.
  http://www.epigenome-noe.net/researchtools/protoco...
  Added: Wed May 27 2009, Hits: 1203, Reviews: 0  Write review   Cached
• Protocol for ChIP-on-chip Experiments (ChIP-on-chip.org)
  This multipart protocol describes procedures for 1) Cross-linking of cells and fragmentation and immunoprecipitation of chromatin; 2) Blunting, ligation of linkers to DNA, and amplification by PCR; 3) Labeling of DNA and microarray hybridization; 4) Microarray analysis and identification of in vivo DNA binding sites
  http://www.chiponchip.org/protocol.html
  Added: Tue Jan 27 2009, Hits: 1615, Reviews: 0  Write review   Cached
• Preparation of Radioactive Probes for Southern Blotting (Gottschling Lab,Fred Hutchinson Cancer Research Center)
  PCR Method to Make Radioactive Probes
  http://labs.fhcrc.org/gottschling/General%20Protoc...
  Added: Sun Nov 01 2009, Hits: 2386, Reviews: 0  Write review   Cached
• Primer-Check (Tiger Team Bioinformatics)
  PrimerCheck displays splice variants and the target location of PCR primers/probes. Optionally it can also display microarray probe target locations.
  http://www.tigerteamconsulting.com/SpliceCenter/Pr...
  Added: Mon Jun 08 2009, Hits: 1969, Reviews: 0  Write review   Cached
• In silico Experiments with Bacterial Genomes (Dr. Joseba Bikandi, University of the Basque Country)
  Theoretical PCR amplification, AFLP-PCR and PFGE with all up-to-date public complete bacterial genomes (139 at the moment). The site is freely available.
  http://www.in-silico.com/
  Added: Thu Oct 30 2003, Hits: 689, Reviews: 0  Write review   Cached
• 3' RACE (Rapid Amplification of cDNA Ends) (Langdale Lab, Department of Plant Sciences, University of Oxford)
  http://dps.plants.ox.ac.uk/langdalelab/protocols/P...
  Added: Wed Feb 11 2009, Hits: 2859, Reviews: 0  Write review  
• Isolation of Retroelement from Plant Genomic DNA (Pat Heslop-Harrison and Trude Schwarzacher, Department of Biology, University of Leicester)
  Retroelements and their derivatives are an ubiquitous and abundant component of plant genomes. From the 1990s, PCR based techniques have been developed to isolate the elements from genomic DNA of different plants, and the methods and primers used are presented here.
  http://www.le.ac.uk/biology/phh4/methods/retros.ht...
  Added: Sat Mar 27 2004, Hits: 74, Reviews: 0  Write review   Cached
• Primer3 (Whitehead Institute/MIT Center for Genome Research)
  Very popular primer design tool for designing primers for PCR, hybridization.
  http://frodo.wi.mit.edu/
  Added: Sat Jul 20 2002, Hits: 8797, Reviews: 0  Write review   Cached
• CODEHOP -COnsensus-DEgenerate Hybrid Oligonucleotide Primers (Blocks WWW Server)
  PCR primers designed from protein multiple sequence alignments
  http://blocks.fhcrc.org/blocks/codehop.html
  Added: Thu Feb 05 2009, Hits: 3012, Reviews: 0  Write review   Cached
• Bisulfite Treatment of DNA (Issa Lab)
  DNA treatment for methylation mapping PCRs such as MSP and bisulfite sequencing PCR
  http://www.mdanderson.org/departments/methylation/...
  Added: Tue May 14 2002, Hits: 2787, Reviews: 0  Write review   Cached
• DNA extraction from Mutation Detection Enhancement (MDE) Gel Stained with Silver Nitrate (Contributed by Mohammad Reza Abbaszadegan et al.)
  Single strand conformational polymorphism (SSCP)is the most widely used PCR-based methods for point mutation detection. The abnormal band found by SSCP analysis is normally verified using sequencing. There are many different procedures to recover DNA fragments from the silver stained polyacrylamide gels. However, most of those techniques are time consuming and their recoveries can be low for the Mutation Detection Enhancement (MDE) gel. This is a rapid and efficient method for recovering DNA fragments from MDE gel. In this procedure, the authors have suggested boiling in phenol/ chloroform/isoamylalcohol for elution of DNA from MDE gel that gives high yield of DNA without swelling of the gel. After precipitation of DNA by ethanol and DNA carrier such as glycogenat -70°C, the DNA pellet is rinsed with 75% ethanol to remove contaminations. The yield and quality of resulting DNA from the SSCP variant by this method is sufficient for PCR re-amplification, which can be followed by DNA sequencing to identify mutation. This method can be similarly used for DNA elution from polyacrylamide gels.
  Added: Wed Feb 19 2003, Reviews: 0  Write review  
• Fungal Genomic DNA Extraction (Contributed by Dr. Eric W. Boehm)
  This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.
  Added: Tue May 14 2002, Reviews: 1 Read reviews  Write review  
• Screen of Recombinant (Chen's Lab)
  Four methods are introduced including clony PCR, boiling ..
  http://www.biochem.ucl.ac.uk/~chen/protocols/colon...
  Added: Tue May 14 2002, Hits: 1575, Reviews: 0  Write review  
• FAQs on Real-Time RT-PCR (Dr. Xiaowei Wang at the Department of Molecular Biology, Massachusetts General Hospital)
  Some questions and answers on real-time PCR primer design and amplification
  http://pga.mgh.harvard.edu/primerbank/faq.html
  Added: Fri May 14 2004, Hits: 1067, Reviews: 0  Write review   Cached
• mRNA Sequencing Sample Preparation Guide (Vincent J. Coates Genomic Sequencing Lab, UC Berkeley)
  This protocol explains how to convert total RNA into a library of template molecules suitable for high throughput DNA sequencing for subsequent cluster generation. The first step in the workflow involves purifying the poly-A containing mRNA molecules using poly-T oligo-attached magnetic beads. Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. This is followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH. These cDNA fragments then go through an end repair process, the addition of a single ‘A’ base, and then ligation of the adapters. These products are then purified and enriched with PCR to create the final cDNA library.
  http://qb3.berkeley.edu/gsl/Protocols_files/mRNA-S...
  Added: Sat Mar 27 2010, Hits: 1113, Reviews: 0  Write review  
• Semi-Quantitative RT-PCR (Mike A. Dyer)
  From RNA isolation, reverse transcription to PCR...
  http://axon.med.harvard.edu/~cepko/protocol/mike/R...
  Added: Tue May 14 2002, Hits: 3161, Reviews: 0  Write review   Cached
• T-Vector Construction (Delaney Lab at the University of Vermont)
  This protocol should work on the basis of the fact that given an equimolar amount of dNTPs in a PCR reaction, Taq DNA polymerase I will add an extra (template-independent) A. If the enzyme is supplied with only dTTP, then an extra T will be added.
  http://www.uvm.edu/~tpdelane/lab/protocols/Tvector...
  Added: Fri Feb 06 2009, Hits: 2659, Reviews: 0  Write review   Cached
• C. elegans microinjection (Dr. Ian Chin-Sang)
  One way to make transgenic animals in C. elegans we use a microinjection technique. Briefly, a DNA construct (plasmid, cosmid or YAC) or PCR product with your gene(s) of interest is mixed with a co-injection marker and injected into the distal gonad (syncytium). The injected DNA is taken up into the mature oocyte's nucleus.
  http://research.abl.es/methods/1441/
  Added: Thu Feb 16 2012, Hits: 914, Reviews: 0  Write review  
• Sequencing of Double Stranded DNA Using Dideoxy Chain Termination (Donis Keller Lab)
  This method is used to sequence double stranded DNA, such as a plasmid insert or purified PCR product. The dideoxy chain termination (or enzymatic) method of DNA sequencing involves the in vitro synthesis of a DNA strand by a modified bacteriophage T7 DNA polymerase (SequenaseR/USB) using a single stranded DNA template.
  http://hg.wustl.edu/hdk_lab_manual/de/de1.html
  Added: Tue May 14 2002, Hits: 1075, Reviews: 0  Write review   Cached

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