Navigation Links
General Notes on Primer Design in PCR*

g said, primers must be chosen so that they have a unique sequence within the template DNA that is to be amplified. A primer designed with a highly repetitive sequence will result in a smear when amplifying genomic DNA. However, the same primer may give a single band if a single clone from a genomic library is amplified.

Because Taq Polymerase is active over a broad range of temperatures, primer extension will occur at the lower temperatures of annealing. If the temperature is too low, non-specific priming may occur which can be extended by the polymerase if there is a short homology at the 3' end. In general, a melting temperature of 55C 72C gives the best results (Note that this corresponds to a primer length of 1824 bases using Wallace's rule above).

Complementary Primer Sequences

Primers need to be designed with absolutely no intra-primer homology beyond 3 base pairs. If a primer has such a region of self-homology, snap back, partially double-stranded structures, can occur which will interfere with annealing to the template.

Another related danger is inter-primer homology. Partial homology in the middle regions of two primers can interfere with hybridization. If the homology should occur at the 3' end of either primer, Primer dimer formation will occur which, more often than not, will prevent the formation of the desired product via competition.

G/C content and Polypyrimidine (T, C) or polypurine (A, G) stretches

The base composition of primers should be between 45% and 55% GC. The primer sequence must be chosen such that there is no PolyG or PolyC stretches tha
'"/>

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:8/26/2014)... 2014 The global market for molecular ... 2020, according to a new study by Grand View ... and the subsequent introduction of advanced cancer diagnostic technologies ... over the next six years. Moreover, the growing global ... cancer, coupled with disease triggering lifestyle habits such as ...
(Date:8/26/2014)... August 26, 2014 Genedata, a ... discovery and life science research, today announced the ... will be held in Asia, Europe, and North ... pharmaceutical companies and leading research organizations will gather ... practices, case studies, and upcoming challenges in all ...
(Date:8/26/2014)... , Aug. 26, 2014   Loveland Products, ... provider of high-performance crop input products and proprietary ... that the company has acquired a controlling interest ... agricultural biotechnology company delivering biochemical-based products for efficient ... Loveland Products, fastest growing plant nutrition brands, Accomplish ...
(Date:8/26/2014)... , Aug. 26, 2014  That,s Nice LLC, a ... today announced the formation of a new dedicated team ... The expanded team will combine current and new staff ... That,s Nice. Guy Tiene ... lead the team. Tiene was previously Director of Marketing ...
Breaking Biology Technology:World Molecular Diagnostics Market Is Expected To Grow To $8,020.1 Million From 2014 To 2020: Grand View Research, Inc 2World Molecular Diagnostics Market Is Expected To Grow To $8,020.1 Million From 2014 To 2020: Grand View Research, Inc 3World Molecular Diagnostics Market Is Expected To Grow To $8,020.1 Million From 2014 To 2020: Grand View Research, Inc 4World Molecular Diagnostics Market Is Expected To Grow To $8,020.1 Million From 2014 To 2020: Grand View Research, Inc 5World Molecular Diagnostics Market Is Expected To Grow To $8,020.1 Million From 2014 To 2020: Grand View Research, Inc 6Thought Leaders to Explore Innovations in SPR & Combination Screening at Genedata Screener User Group Meetings 2Thought Leaders to Explore Innovations in SPR & Combination Screening at Genedata Screener User Group Meetings 3Agrium, through Loveland Products, Acquires a Controlling Interest in Agricultural Biotechnology Company Agricen 2Agrium, through Loveland Products, Acquires a Controlling Interest in Agricultural Biotechnology Company Agricen 3Agrium, through Loveland Products, Acquires a Controlling Interest in Agricultural Biotechnology Company Agricen 4That's Nice Announces Expanded Offering For Strategic Content And Public Relations 2That's Nice Announces Expanded Offering For Strategic Content And Public Relations 3
... Milwaukee Merge eFilm will acquire ... software line in a $6 million cash deal announced ... Merge Technologies Incorporated, expects to close the deal with ... software, used by radiologists and other specialty physicians, includes ...
... are into the home stretch of the year with just December ... market, there are in reality about two weeks left in the ... (as far as IPOs go) by the middle of December. New ... ,What seemed like a promising and valiant biotech IPO market has ...
... in talks with Australian banks , Brookfield, Wis. ... three major Australian banks Commonwealth Bank of Australia, National ... Vipro check processing system. If negotiations are completed successfully, the ... all processing operations managed by Fiserv. , Herrera to join ...
Cached Biology Technology:Midwest life-science company Perrigo globalizes under the radar 2Midwest life-science company Perrigo globalizes under the radar 3Midwest life-science company Perrigo globalizes under the radar 4Midwest life-science company Perrigo globalizes under the radar 5
(Date:8/27/2014)... the brain of mice, scientists have altered the ... by Howard Hughes Medical Institute investigator Susumu Tonegawa ... that the connections between the part of the ... and the part of the brain that stores ... , Altering those connections can transform a ...
(Date:8/27/2014)... Ga. An invasive grass species frequently found in ... who then feed on American toads, a new University ... was accidentally introduced to the U.S. in the early ... and has spread to more than a dozen states ... found along roads and in forests, it can survive ...
(Date:8/27/2014)... CITY, Aug. 27, 2014 University of Utah biologist ... native Turkey, has won that nation,s highest science prize, ... Science. , Sekercioglu is among five researchers picked for ... Technical Research Council of Turkey. Three researchers won the ... the Special Award, which is equivalent to the Science ...
Breaking Biology News(10 mins):Researchers change the emotional association of memories 2Researchers change the emotional association of memories 3Researchers change the emotional association of memories 4More wolf spiders feasting on American toads due to invasive grass, UGA study shows 2More wolf spiders feasting on American toads due to invasive grass, UGA study shows 3University of Utah biologist wins Turkey's top science prize 2
... a switch in cells that may help to kill tumors ... effectively treated by a combination of heat and an anti-cancer ... the enigmatic ability of heat to switch off essential survival ... used, the underlying principles are still unclear. In a ...
... and windy conditions on Black Saturday combined with structures in ... wind flowing through the air - which likely affected fire ... to produce such detailed, high-resolution simulations of weather patterns on ... warning systems. The work was led by Dr Todd ...
... evolution and function of jaws, University of Notre ... His integrative research program investigates major adaptive and ... development and across higher-level groups. In mammals, the ... chewing complex in food procurement and processing has ...
Cached Biology News:Unusual weather events identified during the Black Saturday bushfires 2Notre Dame researcher is shedding light on how jaws evolve 2
... boxes are ideal for use in refrigerators, freezers, or ... Four vibrant colors allow for color coding to project, ... nitrogen drainage or promote cool air circulation. Supplied with ... 100 tubes. Box dimensions: 13 x 13 ...
RABBIT ANTI BACTERIAL HISTIDASE...
REGULATED IN DEVELOPMENT AND DNA DAMAGE RESPONSES 1,HIF-1 responsive RTP801...
Homo sapiens HCLS1 associated protein X-1 Antigen: Recombinant Protein...
Biology Products: