Navigation Links
General Notes on Primer Design in PCR*

Vincent R. Prezioso, PhD
BioSystems Laboratory, Brinkmann Instruments Inc., Westbury, New York

Perhaps the most critical parameter for successful PCR is the design of Primers. All things being equal, a poorly designed primer can result in a PCR reaction that will not work. The primer sequence determines several things such as the length of the product, its melting temperature and ultimately the yield. A poorly designed primer can result in little or no product due to non-specific amplification and/or primer-dimer formation, which can become competitive enough to suppress product formation. This application note is provided to give rules that should be taken into account when designing primers for PCR. More comprehensive coverage of this subject can be found elswhere(1).

Primer selection

Several variables must be taken into account when designing PCR Primers. Among the most critical are:

Primer length
Melting Temperature (Tm)
Specificity
Complementary Primer Sequences
G/C content and Polypyrimidine (T, C) or polypurine (A, G) stretches
3-end Sequence
Each of these critical elements will be discussed in turn.

Primer length

Since both specificity and the temperature and time of annealing are at least partly dependent on primer length, this parameter is critical for successful PCR. In general, oligonucleotides between 18 and 24 bases are extremely sequence specific, provided that the annealing temperature is optimal. Primer length is also proportional to
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. General Considerations for Successful Transfection Experiments
2. Real-Time PCR: General Considerations, Rev A
3. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
4. Eppendorf Electroporation Application Notes Your participation is highly welcome !!!
5. Control RT-PCR Primers for Human Gene Transcripts with Varying Abundance
6. RT-PCR Primer Sets for Human and Mouse Mismatch Repair Genes
7. Gene-Specific Primers for NMDA Receptor Subunits
8. RT-PCR Primers for the Study of Apoptosis
9. New PCR Primers for Human Diseases
10. Amplification of Epstein-Barr Virus Exon C with Difficult Primers Using Eppendorf Mastercycler gradient
11. Human t-PA Control Primer Set
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: General Notes Primer Design PCR

(Date:8/22/2014)... published today (August 22nd), in the scientific journal ... Neuroscience Programme establish the effect of serotonin on ... genetic and optical techniques., "Serotonin is a small ... range of brain functions, from the control of ... emotional behaviours, This neurotransmitter is also popularly thought ...
(Date:8/22/2014)... 22, 2014 Research and Markets ... China Gel Permeation Chromatography (GPC) Industry Report 2014" ... and China Gel Permeation Chromatography (GPC) Industry Report is ... of the global gel permeation chromatography industry with a ... a basic overview of the industry including definitions, classifications, ...
(Date:8/22/2014)... Greenville, SC (PRWEB) August 22, 2014 ... that indisputable acknowledgement is a national disgrace. Furthermore, ... part in establishing the current, deplorable state of ... outlined "5 great mistakes," to merely serve as ... convey the extent of colossal failure(s) that have ...
(Date:8/21/2014)... 2014 2014 Deep Research ... a professional and in-depth research report on ... basic Seaweed Fertilizer information, including Seaweed Fertilizer ... as well as industry overview. This research ... market as well as global industry analysis ...
Breaking Biology Technology:From happiness to pain: Understanding serotonin's function 2Global and China Gel Permeation Chromatography (GPC) Industry Report 2014 25 Great Mistakes in Healthcare and How to Fix Them by Dr. Ira Williams 2Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 2Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 3
... Therapeutics,Inc. (Nasdaq: CVTX ) today reported financial results for ... For the quarter ended December 31, 2007, the Company reported ... compares to a net loss of,$68.1 million, or $1.18 per ... $0.58 per share, for the prior quarter ended September 30,2007. ...
... MANASQUAN, N.J., Feb. 26 BioSpace, the world,s ... career fair company,welcomed over 900 biotechnology and pharmaceutical ... Massachusetts on February 19, 2008., Thirty-six bioscience ... thoroughly impressed with the quality as well as ...
... Microsoft Gold Certified Partner Provides Framework, for ... Solutions, ORLANDO, Fla., Feb. 26 ... software and services for health and,human services ... Certified Partner status in the Microsoft Partner ...
Cached Biology Technology:CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 2CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 3CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 4CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 5CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 6CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 7CV Therapeutics Reports 2007 Fourth Quarter and Full Year Financial Results 8BioSpace Boston Career Fair Attracts over 900 Bioscience Candidates 2Netsmart Technologies Announces Development Program on Microsoft for Local Public Health Clinic Management and Vital Records Software 2Netsmart Technologies Announces Development Program on Microsoft for Local Public Health Clinic Management and Vital Records Software 3
(Date:8/21/2014)... heard of the water window? It consists of radiations ... not absorbed by the water in biological tissues. New ... coherent radiations within the water window. These could be ... high-contrast image of the biological samples or to be ... identifies the physical mechanism needed to efficiently generate the ...
(Date:8/21/2014)... RUSTON, La. A team of researchers ... innovative method for using affordable, consumer-grade 3D ... implants that can contain antibacterial and chemotherapeutic ... team comprised of doctoral students and research ... nanosystems engineering programs collaborated to create filament ...
(Date:8/20/2014)... E. Fox, a John and Rebecca Moores Professor of Biology ... a fellow in the International Society for the Study of ... of four members two from the U.S., one from ... 2014. Fellows are elected every three years, and 36 have ... more than 20 countries, the ISSOL includes researchers from disciplines ...
Breaking Biology News(10 mins):Water window imaging opportunity 2Louisiana Tech University researchers use 3D printers to create custom medical implants 2Louisiana Tech University researchers use 3D printers to create custom medical implants 3UH professor named fellow by International Astrobiology Society 2
... (ESA) will present its fifth annual Regional Policy Award ... during the Society,s upcoming conference in Portland, Oregon. The ... who has an outstanding record of informing political decision-making ... present this prestigious award to Ken Bierly for his ...
... (Boston) A recent study led by Boston University School ... plays a significant role in the regulation of high fat, ... which are published online in PLoS ONE , also ... type 2 diabetes. Katya Ravid, DSc/PhD, professor of ...
... Institution (WHOI) researchers have partnered with two companies ... WHOI: the Imaging FlowCytobot, an automated underwater microscope, ... light to provide wireless transmission of data, including ... WHOI biologists Robert Olson and Heidi Sosik, creators ...
Cached Biology News:Ken Bierly of the Oregon Watershed Enhancement Board to receive ESA Regional Policy Award 2Ken Bierly of the Oregon Watershed Enhancement Board to receive ESA Regional Policy Award 3BUSM study identifies receptor's role in regulating obesity, type 2 diabetes 2WHOI scientists/engineers partner with companies to market revolutionary new instruments 2WHOI scientists/engineers partner with companies to market revolutionary new instruments 3WHOI scientists/engineers partner with companies to market revolutionary new instruments 4
... is pleased to offer the ATTO-TAG ... detection of primary amines. Developed by ... the ATTO-TAG reagents are similar to ... (NDA) and anthracene-2,3-dicarboxaldehyde, all of which ...
...
...
Human Cystatin A MAb (Clone 224705)...
Biology Products: