PCR Protocol--Colony PCR Protocol

Colony PCR Protocols

Colony PCR protocol
Detailed protocol from the web site of the Department of Biology, University of Michigan, USA.www.mcdb.lsa.umich.edu/labs/maddock/protocols/PCR/colony_pcr.html
Colony PCRColony PCR. This protocol is designed to quickly screen for plasmid inserts directly from E. coli. colonies. The plasmid should be high copy number such as ...www.csun.edu/~mls42367/Protocols/Colony%20PCR.pdf
Colony PCR ProtocolColony PCR Protocol. 1. Pull out eight glycerol stock plates from the –80. o. C freezer and set on bench top to. thaw. Be sure to remove the foil seal ...microarrays.berkeley.edu/file_download.php?fid=13
Colony PCR ProtocolColony PCR Protocol. Overnight Culture 5.0µl. 10x PCR buffer 5.0µl. MgCl2 (25mM) 5.0µl. Forward primer (10µM) 0.2µl. Reverse primer (10µM) 0.2µl ...www.unc.edu/~fconlon/Protocols/Colony%20PCR%20Protocol.doc
Colony PCR
Colony PCR Protocol contributed by Lynn Hancock 1. Choose a reasonable sized colony (2-3 mm in diameter) and resuspend 100 µl ddH2O. 2. For PCR, ...www.enterococcus.ouhsc.edu/ColonyPCRProtocol.asp
Yeast Colony PCR -- Amberg et al.
Search CSH Protocols. Advanced Search · Find Protocols · Find a Kit · Access Personal Page · Submit Protocols · Help · Subscribe · About CSH Protocols ...www.cshprotocols.org/cgi/content/short/2006/1/pdb.prot4170
Colony PCR - OpenWetWare
From OpenWetWare. Jump to: navigation, search. See Colony PCR for general information about this protocol and other variants ...openwetware.org/wiki/Endy:Colony_PCR
Colony PCR protocol
Monserate Biotechnology Group Custom services, Colony PCR protocol.www.monseratebiotech.com/colony-pcr.html
Yeast Colony PCR protocolYeast Colony PCR protocol. derived from http://sequence-. www.stanford.edu/group/yeast_deletion_project/deletions3.html.
Colony PCR (Forsburg Lab)
The Forsburg lab pages: fission yeast colony PCR protocol. ... Colony PCR can be used to identify colonies where your favorite gene yfg1 has been replaced ...http://www-rcf.usc.edu/~forsburg/pcr.html
Rapid Screening by Direct Colony PCR Using the FastStart PCR Master
The FastStart PCR Master is a convenient and ideal tool for direct colony PCR. Intact bacteria can be analyzed directly without prior template purification, ...www.analytica-world.com/articles/e/65346/

PCR Protocol--Asymmetric PCR

Asymmetric PCR

What is asymmetric PCR? A PCR in which the predominant product is a single-stranded DNA, as a result of unequal primer concentrations. As asymmetric PCR proceeds, the lower concentration primer is quantitatively incorporated into double-stranded DNA. The higher concentration primer continues to primer synthesis, but only of its strand.

Asymmetric PCR Protocol 1. Pick a phage plaque and place in 100 ul TE or scrape a fresh colony of a bacterial transformant of choice and place in 50 ul of TE/TX100 in a microcentrifuge tube. 2. Heat the tube for 10 min at 95C. 3. Centrifuge at maximum speed for several minutes in a microcentrifuge to pellet cell debris. Collect the supernatant. 4. Add the following components in a PCR tube: 5 ul of phage or bacterial extract (from Step #A3) 50 uM of dNTPs 50 pmol of Primer 1 1 pmol of Primer 2 in 1X PCR Reaction Buffer to give a final reaction volume of 50 to 100 ul 2.5 Units of Taq polymerase 5. Run 30 to 35 cycles in a thermocycler using the following PCR program (see Hint #1 and #2) 95C for 60 sec 60C for 30 sec 72C for 2 min 6. Run a small aliquot on an agarose gel to analyze for single-stranded DNA (see Protocol on Agarose Gel Electrophoresis of DNA). 7. Purify the PCR products and sequence, if desired.
Asymmetric PCR for ssDNA ProductionHOT ASYMMETRIC PCR (Mullins Lab)
Direct sequencing by thermal asymmetric PCR

Rapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers.

Detection of asymmetric PCR products in homogeneoussolution by fluorescence correlation spectroscopy
Asymmetric PCR Using the Primers Anchored on the surface of magnetic nanoparticles.

PCR Protocol--Long PCR Protocol

Long PCR Protocol

Kyle’s protocol for long PCR
Kyle’s protocol for long PCR. This protocol intended for generation of long PCR products (3-10kb) from mouse ES cell or tail DNA. General Considerations: ...www.whitelabs.org/Lab%20Protocols/PCR%20Protocols/kyle.htm
Long PCR Protocol
Protocol and guidelines for choice of conditions for PCR of long sequences (10 kb or larger). From Genetics Dept., Harvard Medical School, Boston, MA, USA.arep.med.harvard.edu/labgc/estep/longPCR_protocol.html
Tips for Long and Accurate PCR Tips for Long and Accurate PCR. Wayne M. Barnes. Department of Biochemistry and Molecular Biophysics, Washington University School of ...http://www.klentaq.com/products/lapcrchapter.pdf
Mullins Lab : Protocols : Long PCR
Two long PCR steps:. First round of the nested PCR step of the end point dilution procedure to quantitate the cDNA. First round of the nested PCR step of ...ubik.microbiol.washington.edu/protocols/other/longpcr.htm
Long PCR
LONG PCR AMPLIFICATION OF THE FVIII GENE INTRON 22 GENE INVERSION. Introduction. Long range PCR allows the amplification of PCR products, which are much ...europium.csc.mrc.ac.uk/WebPages/Database/Methods/longpcr.htm
Extra-Long PCR Ki, EPICENTRE Biotechnologies
EPICENTRE develops and manufactures reagents and kits for genomics, proteomics, and RNA research. EPICENTRE's products are used worldwide in a broad range ...www.epibio.com/item.asp?ID=306
Long PCR Protocol
Efficient Long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading ...wheat.pw.usda.gov/~lazo/methods/lazo/longpcr.html
Long PCR.
Long PCR. Cheng S, Chang SY, Gravitt P, Respess R. Department of Human Genetics, Roche Molecular Systems, Inc., Alameda, California 94501. Mesh Terms: ...www.ncbi.nlm.nih.gov/pubmed/8208299
Long PCR Product Sequencing (LoPPS): a shotgun-based approach to sequence long PCR products.
Here we describe a practical procedure for sequencing long PCR products. The method relies on ultrasonic shearing of PCR products, resulting in fragments ...www.nature.com/nprot/journal/v2/n2/abs/nprot.2006.453.html
Long PCR Protocol
Aug 10, 2003 ... Introduction · Research Pages · People involved Photo galleries Publications Other sample sites Education · Links to other micobial ...www.hawaii.edu/microbiology/MO/longpcr.htm
Long PCR -- Sambrook and Russell.
Primers used for long PCR are generally slightly longer (25-30 nucleotides) than those used for standard PCR. It is particularly important to strive for ...www.cshprotocols.org/cgi/content/full/2006/2/pdb.prot3841
Efficient Cloning of Long PCR Inserts with the In-Fusion PCR Cloning System.Clontech’s In-Fusion PCR cloning method. provides superior results when applied to the. challenging task of cloning long PCR fragments. ...www.clontech.com/images/ctq/APR07UPD/CR742319_LongPCR_IF_US.pdf
Nature Protocols: Long PCR amplification of the entire mitochondrial genome from individual helminths for direct sequencing.
In this article, we describe a practical long PCR approach for the amplification and subsequent sequencing of the entire mt genome from individual helminths ...www.natureprotocols.com/2007/09/20/long_pcr_amplification_of_the.php
Long PCR - MyBio
Long PCR Reagents and GuidelinesLong PCR Reagents and Guidelines. ... General Guidelines for Long PCR Conditions and Enzyme Mixtures Efficient Long PCR ...mybio.wikia.com/wiki/Long_PCR

PCR Protocol--Methylation Specific PCR

Methylation-specific PCR: a novel PCR assay for methylation status ...
Selected References. Page Browse · PDF (1.9M) · Contents · Archive. Related material:. PubMed record, PubMed related arts, PubMed LinkOut, Substance ...www.pubmedcentral.nih.gov/articlerender.fcgi?artid=38513

PCR Protocol--PCR Application Manual

PCR Applications Manual (from Roche Diagnostics).

PCR Protocol--RAPD PCR

RAPD PCR

RAPD PCR
RAPD stands for Random Amplification of Polymorphic DNA, where the target sequence(s) (to be amplified) is unknown.Brief description, from Rutgers ...avery.rutgers.edu/WSSP/StudentScholars/project/archives/onions/rapd.html
RAPD PCR Reaction Mixes:
Type, Initialize, X, Denature, Anneal, Extend, Termination. RAPD 10-mer, 95C, 5min, 45, 94C, 0:30, 35C, 1:00, 72C, 2:00, 72C, 5:00. PCR 18+mer, 95C, 5:00 ...wheat.pw.usda.gov/~lazo/methods/lazo/pcrmeth0.html
Handbook for DNA isolation, RAPD-PCR and PCR-RFLP
A standard RAPD-PCR reaction is performed in volumes of 25.0 m l containing 50 mM KCl, 10 mM Tris-HCl pH 9.0, 1.5 mM MgCl2, 0.001% gelatine, ...www.toyen.uio.no/botanisk/brochmann/handbook.htm
RAPD Protocol dNTP mix (5mM): Loading buffer (6X): Ladder mix: PCR .RAPD Protocol. Protocol for Promega. and Stoffel brand Taq. enzymes (1 reaction):. 4 µl dNTP (5mM) mix. 2.5 µl 10X PCR buffer. 8.33 µl H ...www.css.msu.edu/bean/PDF/RAPD_protocol.pdf
PCR PROTOCOL FOR RAPDs--1PCR PROTOCOL FOR RAPDs. Application: Fingerprinting, molecular mapping, ... Amount of DNA needed per RAPD reaction according to the plant species under ...gene4.ciat.cgiar.org/limsys3.0/Modelos/protocolos/PCRPROTOCOLFORRAPDs.doc

PCR Protocol--RACE PCR

RACE PCR

Background: Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction-
based technique which facilitates the cloning of full-length cDNA
sequences when only a partial cDNA sequence is available. Traditionally,
cDNA sequence is obtained from clones isolated from plasmid or
phage libraries. Frequently these clones lack sequences corresponding to
the 5' ends of the mRNA transcripts. The missing sequence information
is typically sought by repeatedly screening the cDNA library in an effort
to obtain clones that extended further towards the 5' end of the message.
The nature of the enzymatic reactions employed to produce cDNA
libraries limits the probability of retrieving extreme 5' sequence even
from libraries that are very high quality.

RACE-PCR facilitates the isolation of 5 and 3 end sequences from cDNA.
Human Molecular Genetics 2 conveys the feel of fast-moving research while providing a description in some depth of the techniques and data that are helping ...www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=hmg.figgrp.2574
5' RACE PCR
5' RACE PCR Protocol 1. First strand cDNA synthesis: - Reaction set up: cDNA synthesis buffer 4 ul dNTP mixture 2 ul gene specific primer 1 1 ul ...biowww.net/detail-126.html
3' RACE PCR
Detailed protocol on 3' RACE PCR - rapid amplification of 3' cDNA end. ... 3' RACE PCR Protocol: 1. First strand cDNA synthesis: ...biowww.net/detail-329.html SMART RACE cDNA Amplification Protocol-at-a-GlanceProtocol-at-a-Glance. 2. For 5'-RACE and 3'-RACE: prepare PCR reactions as shown in Tables III & IV. Add the components in the order shown in PCR tubes. ...www.clontech.com/images/pt/PT3269-2.pdf
5' RACE Protocol For Generation of Seq. Tags.Apply all 79 ul of1st PCR product to center of packed wells. ... Sequence template per 5'RACE sequencing protocol with sequencing oligo 305. ...baygenomics.ucsf.edu/protocols/comp1/RACE_generation.pdf
Rapid Amplification of cDNA 3' Ends (3'- RACE)( Simplified protocol based on Clontech’s SMART- RACE ). 6. Distribute 5 µl of diluted first PCR product in the PCR tubes, 8. strips or 96 plate on ice. ...www.cmhd.ca/protocols/genetrap_pdf/3'RACE.pdf
A lock-docking oligo(dT) primer for 5' and 3' RACE PCR.
stretch of As, yielding a discrete PCR. product. We are planning to use the lock-dock-. ing primer in the 5'-RACE protocol (1) to ...www.genome.org/cgi/reprint/2/2/144.pdf
First Choice RACE-Ready cDNA ProtocolThe 3' RACE protocol. describes nested PCR, however 3' RACE reactions may produce signif- ..... your PCR protocol. Figure 3. 3' RACE Control. 3' Adapter ...http://www.ambion.com/techlib/prot/fm_3200.pdf
Nature Protocols: 3′ End cDNA amplification using classic RACE
... amplification of cDNA ends (RACE) PCR as long as part of the mRNA sequence is known; ... cDNA ends can be obtained in 1–3 days using this protocol. ...www.natureprotocols.com/2007/01/11/3_end_cdna_amplification_using.php
High throughput Rapid Amplification of cDNA Ends (RACE) using cDNA Library as Template.Follow standard protocol for plasmid prep in microtitterplates. PCR for RACE-clones:. Use standard protocol for PCR, but use 1 ul plasmidprep as template ...www.bio.aau.dk/download/biotechnology/kln/RACE-procedures.doc
Help for 3'-RACE PCR
Help for 3'-RACE PCR. PROBLEM: We have prolem to amplify the 3'-end of our .... I intend to try the following: a) Modified an inverse PCR protocol by ...
http://wheat.pw.usda.gov/~lazo/methods/lazo/met11.html