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:11/21/2014)... Author Matthew J. Pallamary’s second short ... Side” published as a tribute to his mentor ... Short Story” category of the 2014 USA Best ... of USA Book News, said this year’s contest yielded ... Simon & Schuster, Penguin, John Wiley & Sons, Houghton ...
(Date:11/21/2014)... November 20, 2014 The November 2014 ... as an open access journal featuring the following articles:, ... proteins to preserve photoresponsiveness in the retina , ... in situ hybridization using chromogenic substrates in zebrafish , ... RNA, and protein electrophoresis , Nuclear LC3-positive ...
(Date:11/21/2014)... 2014 Deep Research Report on ... professional and in-depth research report on the Sodium ... like its definition, classification, application, and industry chain ... covers the international market analysis, including China’s domestic ... US, Europe, Asia, China, Japan etc. regions) industry ...
(Date:11/21/2014)... - Aequus Pharmaceuticals Inc. (the "Company" or "Aequus") ... a brokered private placement offering (the "Brokered Offering") ... Securities Inc. and Clarus Securities Inc. acted as ... syndicate of agents that also included Wolverton Securities ... Concurrently with the Brokered Offering, the Company also ...
Breaking Biology Technology:Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 2Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 3Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 4BioTechniques: The International Journal of Life Science Methods Announces the Publication of the November 2014 Issue—Available in Print and Online 2BioTechniques: The International Journal of Life Science Methods Announces the Publication of the November 2014 Issue—Available in Print and Online 3Sodium Ferrocyanide Market: Analysis for Global & China Regions Development Trends Available at DeepResearchReports.com 2Sodium Ferrocyanide Market: Analysis for Global & China Regions Development Trends Available at DeepResearchReports.com 3Aequus Pharmaceuticals closes $4.2 million private placement financing 2Aequus Pharmaceuticals closes $4.2 million private placement financing 3
... (NTI (R)) (Nasdaq: NTII ) today announced the ... 31, 2007., Paul Freiman, president and chief executive ... occasion of his 75th birthday. Dr.,Cape is a legend ... board,has been outstanding. Prior to joining the board, Dr. ...
... FRANCISCO, Calif., and GREENWICH, Conn., Dec. 20 /PRNewswire-,FirstCall/ ... and GBP IP, LLC, an,affiliate of GBP Capital, ... announced that the two companies have entered,into an ... gene,delivery technology, commonly referred to as lentiviral vectors. ...
... 20 Memory,Pharmaceuticals Corp. (Nasdaq: MEMY ) ... randomized, double-blind, placebo-controlled Phase 2a clinical,trial of MEM ... in cognitive impairment associated with schizophrenia (CIAS). The,trial ... cognitive effects,of three doses of MEM 3454 in ...
Cached Biology Technology:Cell Genesys Announces Sale of Lentiviral Gene Delivery Technology for $12 Million 2Cell Genesys Announces Sale of Lentiviral Gene Delivery Technology for $12 Million 3Cell Genesys Announces Sale of Lentiviral Gene Delivery Technology for $12 Million 4Cell Genesys Announces Sale of Lentiviral Gene Delivery Technology for $12 Million 5Memory Pharmaceuticals Initiates Phase 2a Trial of MEM 3454 in Cognitive Impairment Associated with Schizophrenia 2Memory Pharmaceuticals Initiates Phase 2a Trial of MEM 3454 in Cognitive Impairment Associated with Schizophrenia 3Memory Pharmaceuticals Initiates Phase 2a Trial of MEM 3454 in Cognitive Impairment Associated with Schizophrenia 4
(Date:11/21/2014)... JOSE, Calif. , Nov. 20, 2014   ... global leader in microcontroller (MCU) and touch technology solutions, ... digital temperature sensors with the widest V cc ... delivers higher temperature accuracy and faster I 2 C ... registers and serial EEPROM memory making them ideal for ...
(Date:11/18/2014)... --  News Highlights: , ... Partners Data Lake, an agile data and analytics ... allow researcher and clinicians to explore and develop ... of patients , The Partners Data Lake ... Partners system, breaking down physical barriers for collaboration ...
(Date:11/18/2014)... NEW YORK , Nov. 18, 2014 /PRNewswire/ ... iris-based identity authentication solutions, and MorphoTrust USA ... solutions and services, today announced a strategic partnership ... across enterprise, motor vehicle administration (MVA), airport screening ... leader in identity-related solutions, MorphoTrust serves consumers through ...
Breaking Biology News(10 mins):Atmel Launches Industry's First Wide-V(cc) Low-Power Temperature Sensor Family 2Atmel Launches Industry's First Wide-V(cc) Low-Power Temperature Sensor Family 3Atmel Launches Industry's First Wide-V(cc) Low-Power Temperature Sensor Family 4Partners HealthCare and EMC Unite to Solve Health Care's Toughest Challenges Using Big Data 2Partners HealthCare and EMC Unite to Solve Health Care's Toughest Challenges Using Big Data 3Partners HealthCare and EMC Unite to Solve Health Care's Toughest Challenges Using Big Data 4Partners HealthCare and EMC Unite to Solve Health Care's Toughest Challenges Using Big Data 5EyeLock, MorphoTrust Form Strategic Partnership to Pursue Commercial Enterprise Deployments 2EyeLock, MorphoTrust Form Strategic Partnership to Pursue Commercial Enterprise Deployments 3
... , Results from the first ever European-wide retrospective analysis ... both D-penicillamine and trientine continue to be effective treatments, ... free from a liver transplant. , Wilson disease is ... not excreted by the body effectively, leading to excess ...
... Researchers at The Johns Hopkins University School of ... to become contaminated with unacceptably high levels of bacteria, ... operated faucets. The study will be presented on ... Healthcare Epidemiology of America (SHEA). Electronic-eye, non-touch faucets ...
... Va.--The Department of the Navy announced today the award ... academic institutions as part of the Office of Naval ... Young Investigator Program is an important part of the ... (STEM) outreach programs," said Secretary of the Navy Ray ...
Cached Biology News:European-wide study confirms benefits of D-penicillamine and trientine for Wilson disease 2Electronic faucets unsafe for use in high-risk patient hospital settings 2
... have been developed for affinity capture ... variety of ProteinChip chemical surfaces allow ... their unique biochemical properties. Each chip ... sample comparison, and the chips are ...
... been developed for affinity capture of ... of ProteinChip chemical surfaces allow differential ... unique biochemical properties. Each chip has ... comparison, and the chips are color-coded ...
Freeze 'N Squeeze DNA gel extraction spin columns use a quick freeze-and-thaw cycle and centrifugation to draw 50-23,000 bp DNA out of agarose gel slices. Supplied as 25 columns....
The Freeze 'N Squeeze DNA gel extraction kit uses a quick freeze-and-thaw cycle and centrifugation to draw 50-23,000 bp DNA out of agarose gel slices. The kit includes materials for 100 preps....
Biology Products: