Navigation Links
Using Gradient PCR to determine the optimum annealing temperature.

Problems with the amplification of a specific DNA fragment using the PCR* are an everyday occurrence in the lab. For example, unspecific secondary bands may form after the PCR reaction, which hinder, or even prevent, further analyses (cycle sequencing, mutation detection, etc.) or an unequivocal assessment of the PCR result. In such cases, PCR conditions must be optimized. This is normally achieved by titrating the magnesium-, template-, primer-, dNTP- and Taq polymerase concentration, Hot Start PCR, Touch-down PCR, adding detergents, reducing the PCR cycles or by gradually increasing the annealing temperature. Fig. 1: Experimental determinati on of optimal annealing temperature: The calculated primer annealing temperature was 56.5C, the actual annealing temperature is 63.5C. The ribosomal spacer region of mycoplasms from H9 cell cultures was amplified. Using the gradient function of the universal block, a gradient of 53 to 67C was set. The following test parameters were selected: denaturation 10 s, annealing 15 s, elongation 20 s, amount of Taq-polymerase 0.75 units; duration of entire experiment: <1 h. Fig. 2: Amplification shown in fig. 1 performed under optimized temperature conditions. In this experiment, the universal block was set to a uniform temperature in the annealing phase. The outstanding temperature homogeneity of the block ensures reproducible PCR results. The selection of the annealing temperature is possibly the most critical component for optimizing the spec
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
2. Oil-Free PCR Using the Hot Top Assembly
3. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
4. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
5. Using XL1-Red Mutator Strain to Generate Esterase Variants
6. Detect Released Oligosaccharides Using the Eagle Eye II Still Video Imaging System
7. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
8. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
9. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
10. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
11. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Using Gradient PCR determine the optimum annealing temperature

(Date:8/31/2015)... ON , Aug. 31, 2015 /PRNewswire/ - BIOREM Inc. ... orders totalling $5.7 million. The orders are for air emission abatement ... "Of particular note is a large order for a municipality ... equipment will be replacing a competitor,s system that failed to ... in our engineered solutions," said Derek S. Webb , ...
(Date:8/31/2015)... Aug. 31, 2015 Research and Markets ( ... "Israel Biomedical Sensors Market - Growth, Trends & Forecasts ... The Israel Biomedical Sensors market is estimated at $0.19 ... the period 2014-2020 The near future will ... formulation of each individual. These sensors can be set ...
(Date:8/31/2015)... SYDNEY , Aug. 31, 2015  US-Australian ... NVGN ) today confirmed its comprehensive scientific ... ground-breaking technology platforms in areas of unmet patient ... through the remaining preclinical projects prior to entering ... lead compounds. Acting Chief Executive Officer, ...
(Date:8/31/2015)... ... 31, 2015 , ... Regulatory agencies have acknowledged impurities especially ... process. Thus, innovator companies must understand what will be needed in the drug ... been hosting multiple educational panels and seminars for experts to share advice concerning ...
Breaking Biology Technology:Israel Biomedical Sensors Market 2015 - Growth, Trends & Forecasts to 2020 2Novogen Announces The Outcome Of A Comprehensive Science Review 2Novogen Announces The Outcome Of A Comprehensive Science Review 3Novogen Announces The Outcome Of A Comprehensive Science Review 4Novogen Announces The Outcome Of A Comprehensive Science Review 5Novogen Announces The Outcome Of A Comprehensive Science Review 6Regis Shares Expert Advice on Impurities in Drug Development 2Regis Shares Expert Advice on Impurities in Drug Development 3
... Researchers from A*STAR Singapore took lead roles in a ... during long-term culture of human embryonic stem cells (hESCs). ... Peter Andrews of the University of Sheffield (UK), Paul ... Oh of Singapore,s Bioprocessing Technology Institute (BTI), and Barbara ...
... November 28, 2011 Elsevier ... analytic solutions, today announced that Thomas H. Zajac ... / MEDai provides solutions for providers, health plans ... profiling, regulatory compliance and clinical surveillance to improve ...
... Pelleve™ Wrinkle Reduction System was featured in a live ... Top dermatologist Dr. Ava Shamban of Santa Monica, CA ... first-hand look at how Pelleve works. The treatment produced ... takes to watch the show. A revolutionary, ...
Cached Biology Technology:Singapore scientists lead human embryonic stem cell study 2Singapore scientists lead human embryonic stem cell study 3Elsevier / MEDai Appoints Tom Zajac as New President 2Elsevier / MEDai Appoints Tom Zajac as New President 3Ellman's Pelleve™ Wrinkle Reduction System featured in "Sexy Eyes" segment on The Doctors 2
(Date:8/3/2015)... Synaptics, Inc. (Nasdaq: SYNA ), the ... members of the executive management team will present to ... Leadership Forum on Tuesday, August 11, 2015 at 1:00 ... the Sonnenalp Resort in Vail, CO. ... information. An audio webcast of the event will be ...
(Date:7/31/2015)... , 31 de julio de 2015 BGI llevará ... www.icg-10.org ) del 22 al 25 de octubre de ... La conferencia celebra su décimo aniversario este año. ... en una de las reuniones anuales más influyentes del ... una de las conferencias científicas más dinámicas, entusiastas y ...
(Date:7/31/2015)... , Chine, 31 juillet 2015 ... www.icg-10.org ) sera organisée par le BGI ... Shenzhen en Chine. ... Depuis son inauguration en 2006, l,ICG est devenue ... dans le domaine des « omiques » et c,est aussi ...
Breaking Biology News(10 mins):La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 2La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 3La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 2La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 3
... part of the Genome Canada Large-Scale Applied Research Project ... to 16 projects across the country, the researchers in ... with the aim of identifying genes or proteins for ... target cancers and other diseases. The first project, ...
... - New research from the University of Calgary,s Faculty ... diagnosis using an iPhone application with the same accuracy ... technology can be particularly useful in rural medical settings. ... neurologists, regardless of where the physicians and patients are ...
... has revealed how infection by a parasitic fungus dramatically changes ... leonardi ), causing them to become zombie-like and to die ... fungus. The multinational research team studied ants living high up ... research will be published in the BioMed Central open-access journal ...
Cached Biology News:OGI genomics researchers awarded $23 million 2OGI genomics researchers awarded $23 million 3iPhone can diagnose stroke: study 2Zombie ants have fungus on the brain, new research reveals 2
... random mutations to the gene supplied by a ... a vector of choice to obtain a library ... be set to a desired value within 1-20 ... Mutations can be restricted to a desired ...
... plasmid DNA preparations are prepared from ... from 1mg to multi-gram scales. Each ... needs, and glycerol stocks are maintained ... clonal selection (Clonal selection includes screening ...
... sports 16-independent channels for automated gigaseal patch ... screening against voltage-gated ion channels, such as ... channels. The four pipette heads afford ... ion channel drug discovery. The glass-coated ...
... to be able to handle larger sample volumes, ... well as interface with telemetry systems. The ... with dogs, pigs, and primates. The system ... and the computer can be located outside the ...
Biology Products: