Navigation Links
Real-Time PCR: General Considerations, Rev A

mized primer set for a quantitative real-time assay.


Considerations for General PCR Optimization
General Laboratory Practices for Quantitative Real-Time PCR
In general, follow these practices to ensure the highest probability of success:
Wear gloves
Use screwcap tubes
Use aerosol-resistant filter tips
Use calibrated pipets dedicated to PCR
Use PCR-grade water and use only for PCR
Use a no-template control to verify absence of contamination
Prepare reactions in replicate ideally as triplicates

Replicate Quality
To obtain good replicates, a master mix should be prepared with all reaction components including the sample Use a hot-start enzyme to prevent nonspecific amplification during preparation
Make up a master mix with sufficient volume to prepare all replicate samples
Pipet once per well


DNA Source
The source of the template affects the accessibility of the target sequence and must be considered during optimization. It is important to optimize the reaction for the template concentrations that will be used in your experiment.

Genomic DNA (Intact, High Molecular Weight DNA)
Cut with a restriction enzyme that does not cut within the region to be amplified
Boil DNA stock for 10 min and place immediately on ice

Plasmid DNA
If there are problems with amplification, linearize the plasmid with a restriction enzyme that does not cut within the target

cDNA
RNA must be free from genomic DNA contamination treat with RNase-free DNase prior to reverse tr
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
2. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
3. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
4. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
5. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
6. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
7. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
8. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
9. Real-Time Immuno-PCR on the iCycler iQ System, Rev A
10. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
11. General Notes on Primer Design in PCR*
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Real Time PCR General Considerations Rev

(Date:2/27/2015)... BERKELEY, Calif. , Feb. 27, 2015  Steep ... in the US and internationally, intends to open a ... Albuquerque, New Mexico , bringing advanced scientific ... of cannabis. Steep Hill is currently the ... Health to conduct potency and contaminant testing in order ...
(Date:2/26/2015)... Feb. 26, 2015  23andMe, the leading personal genetics ... as Privacy Officer and Corporate Counsel. Black brings ... laws as well as health care regulations. As a ... be responsible for reviewing, updating and enhancing the company,s ... and abroad. She joined the company January 5, 2015 ...
(Date:2/26/2015)... Protiviti ( www.protiviti.com ), a global consulting ... Governance Portal , designed to increase the efficiency with ... across multiple business units and projects. The Governance Portal ... align their controls with the COSO ( Committee of ... response to client feedback, Protiviti has also included a ...
(Date:2/26/2015)... , Feb. 26, 2015  NuGene International, ... aesthetic products for skin and hair rejuvenation, has ... as agency of record for investor relations and ... improve our distribution, sales and diversification strategies – ... current and prospective shareholders," stated Ali Kharazmi CEO ...
Breaking Biology Technology:Steep Hill First to Receive Provisional Approval to Establish Laboratory Services for Cannabis Testing in State of New Mexico 223andMe Appoints Kate Black as Privacy Officer and Corporate Counsel 2Protiviti Governance Portal Helps Companies Drive Governance, Risk and Compliance Productivity Across Multiple Business Units 2Protiviti Governance Portal Helps Companies Drive Governance, Risk and Compliance Productivity Across Multiple Business Units 3NuGene International, Inc. Selects Capital Market Relations for Strategic Investor Relations 2NuGene International, Inc. Selects Capital Market Relations for Strategic Investor Relations 3
... N.J., Sept. 2 PTC Therapeutics, Inc. (PTC) today announced ... advancing drug discovery efforts on an oncology target addressed through ... technology. , , (Logo: ... In September 2007, Celgene made a $20 million equity investment ...
... , SOUTH PLAINFIELD, N.J., Sept. 2 ... research collaboration and licensing agreement with Roche for the development of ... by Small-molecules (GEMS(TM)). The collaboration focuses initially on four CNS ... (Logo: http://www.newscom.com/cgi-bin/prnh/20010919/PTCLOGO ) , ...
... Predatory fish are well aware of the problem: In a ... A similar situation can be found in the microcosm of ... electrons. In order to achieve control over single electrons in ... are needed. Physicists of the Max Planck Institute of Quantum ...
Cached Biology Technology:PTC Therapeutics Announces Celgene Exercised Option to Drug Discovery Collaboration 2PTC Therapeutics Announces Drug Discovery Collaboration With Roche 2PTC Therapeutics Announces Drug Discovery Collaboration With Roche 3Laser pulses control single electrons in complex molecules 2Laser pulses control single electrons in complex molecules 3
(Date:2/5/2015)... 2015 Despite a lack of advancements ... as consumer electronics, automotive, storage and telecommunication has ... (SMT) screen printers. Innovations in electronics and conversion ... the adoption curve up. Meanwhile, sale volumes of ... dispensers are indispensable in the production of complex ...
(Date:1/22/2015)... 13, 2015  Today, FreeWavz ( www.FreeWavz.com ), the ... campaign on Fundable, https://www.fundable.com/freewavz . FreeWavz will ... capacity to meet customer demand. Logo ... http://photos.prnewswire.com/prnh/20150113/168790 Invented by Dr. ...
(Date:1/22/2015)... OXFORD, Conn. , Jan. 20, 2015 NXT-ID, Inc. (NASDAQ: ... growing mobile commerce market, reports on the recent success of the Wocket™ ... Wocket smart wallet was named as one of the ... of the "5 Best Products Launched At CES So Far" by Newseveryday.com ...
Breaking Biology News(10 mins):End-user Demand Spurs Global SMT Screen Printers and Glue Dispensers Market 2End-user Demand Spurs Global SMT Screen Printers and Glue Dispensers Market 3FreeWavz Launches on Fundable to Drive Speed-to-Market in 2015 2CES Response for NXT-ID's Wocket Smart Wallet Kicks Off 2015 2CES Response for NXT-ID's Wocket Smart Wallet Kicks Off 2015 3
... brake linings and mining operations can affect salmon to ... Washington State University researcher. Jenifer McIntyre found the metal ... won,t detect a compound that ordinarily alerts them to ... not getting the information it needs to make good ...
... -- Like their warrior ancestors, leaders of many Pacific Island ... time by sounding an alarm as the impact of climate ... big boost with the release of a Scientific Consensus Statement ... over 2,400 scientists, showing the threats that reef corals are ...
... computer one recent afternoon, University at Buffalo researcher Charmion Cruickshank ... chemicals in the urine of a child with autism. ... with the disorder and five without -- and she has ... data for autism,s chemical clues. The goal of the ...
Cached Biology News:Copper making salmon prone to predators 2Scientists join forces in call for action to save coral reefs 2Scientists join forces in call for action to save coral reefs 3Hunting for autism's chemical clues 2