Navigation Links
Amplification of Epstein-Barr Virus Exon C with Difficult Primers ,,, Using Eppendorf Mastercycler gradient

g reactions. For this reason, primers flanking exactly the 5'-terminal and the 3'-terminal of each exon were synthesized in order that the cloned amplimer will be in-framed during subsequent clonings (Fig.1). In so doing, we were restricted to only one choice for each primer. Exons A and B were successfully amplified by their respective primers at the calculated annealing temperature using a conventional thermal cycler. However, this was not the case for Exon C. The properties of primers used for amplifying Exon C are tabulated in Table 1.



Table 1: Properties of primer CF and CR specific for Exon C of LMP-1 Methods

During optimization for primers CF and CR, we applied the general rule of starting with annealing temperatures around 5C below the calculated melting temperature (Tm ) and titrating for optimal MgCl2 concentrations. The Tm of both primers were recommended by the manufacturer (Operon, Inc., Alameda, CA, USA) calculated using the standard formula. Using a conventional thermal cycler, we were unable to amplify Exon C despite attempts over a range of annealing temperatures (52C to 65C) and MgCl2 concentrations (1 mM to 4 mM). This problem was solved when we used the temperature gradient function of the Eppendorf Mastercycler gradient.4,5

By performing a wild search for the correct annealing temperature, we subjected our amplification reaction using primers CF and CR (Table 1) over a temperature gradient of 20C (48C to 68C) at a constant MgCl2 concentration of 2 mM. Template EBV DNA was extracted from B95.8 marmoset lymphocytes cell line. PCR was carried out in 50 l reactions with the f
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Greater Amplification Specificity with New Hot Start PCR Enzyme
4. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
5. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
6. Improve Amplification Specificity with Hot Start PCR Enzyme
7. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
8. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
9. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
10. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
11. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
Post Your Comments:
(Date:9/19/2014)... yields to disease risks, the biological characteristics people care ... as for heightthe textbook example of a complex traitattributes ... by multiple genetic and environmental influences, making it challenging ... genes, geneticists typically mate two individuals that differ in ... mouseand then study their descendents, looking for genes that ...
(Date:9/19/2014)... Biotech companies make announcements illustrating ... systems and advanced Laboratory Instrumentation. Companies in focus today ... BDX ), Thermo Fisher Scientific Inc. (NYSE: ... RGDO ), Sangamo Biosciences Inc. (NASDAQ: SGMO ... BioSciences, Inc. (OTCQB: PBIO) ("PBI" or the "Company") today ...
(Date:9/19/2014)... (PRWEB) September 19, 2014 Cereal ... food industries will gather October 5–8, 2014, for ... Rhode Island Convention Center in Providence, Rhode Island. ... the work of food scientists, chemists, microbiologists, nutritionists, ... and food production management. To date, 1,000 attendees ...
(Date:9/18/2014)... HealthTronics, Inc ., a leading ... and services, announced that a poster related to ... external beam radiotherapy (EBRT) is being presented at ... (MAAUA) Annual Meeting in Baltimore, Maryland September 18-21, ... and External Beam Radiation as Treatments for Localized ...
Breaking Biology Technology:Want to link genes to complex traits? Start with more diversity 2Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 2Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 3Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 4Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 5Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 6Medical Instrumentation Automation Advances for Biological Sample Preparation - Company Expects Q4 Revenue Impact from Newest Instrument Systems 7Latest Research on Cereal Grain Quality, Health Benefits and Food Production to be Presented During the AACC International Annual Meeting 2Latest Research on Cereal Grain Quality, Health Benefits and Food Production to be Presented During the AACC International Annual Meeting 3HealthTronics Announces Data Comparing Cryotherapy Versus Radiation for Treating Prostate Cancer Poster Presented at the American Urological Association Regional Meeting 2
... , , LONDON and ... Silence Therapeutics plc (AIM: SLN) today announced that Philip Haworth ... update at the 12th Annual BIO CEO & Investor Conference.  The conference ... in New York City . , Silence,s presentation will take place ...
... OAK RIDGE, Tenn., Feb. 4, 2010 -- Four Oak ... that will receive five-year research grants as part of ... The $85 million program, funded under the American ... Science, is designed to support exceptional researchers during the ...
... , Feb. 4 A comprehensive study of ... community oncology practices revealed that Medicare covers only 56% of ... room services to seniors with cancer. The remaining costs -- ... and follow-up care planning -- are not reimbursed by Medicare, ...
Cached Biology Technology:Silence Therapeutics to Present at 12th Annual BIO CEO & Investor Conference 2Silence Therapeutics to Present at 12th Annual BIO CEO & Investor Conference 34 ORNL researchers selected for Recovery Act early career funds 24 ORNL researchers selected for Recovery Act early career funds 3Study: Medicare Covers Only Half the Cost of Administering Chemotherapy to Seniors with Cancer 2Study: Medicare Covers Only Half the Cost of Administering Chemotherapy to Seniors with Cancer 3Study: Medicare Covers Only Half the Cost of Administering Chemotherapy to Seniors with Cancer 4
(Date:9/18/2014)... new GSA Bulletin study uses tree rings ... and Chaco Wash in northern New Mexico, USA. By ... and willow, investigators were able to precisely date arroyo ... combined this data with aerial imagery, LiDAR, longitudinal profiles, ... arroyos. , Arroyos are deep, oversized channels that have ...
(Date:9/18/2014)... and the Quebec government have discovered microplastics (in ... of Fisheries and Aquatic Sciences . , The ... industrial cleansers, to which they are commonly added ... buoyancy, they may readily pass through sewage treatment ... the world,s oceans, but have only recently been ...
(Date:9/18/2014)... DNA, tiny strands of nucleic acid that contain instructions ... genetic data, our DNA is copied into RNA molecules, ... tasks in our cells. , Several years ago, ... all other known RNAs, this molecule is circular, and ... little has been known about how they are produced. ...
Breaking Biology News(10 mins):Tree rings and arroyos 2Tree rings and arroyos 3Tree rings and arroyos 4Tree rings and arroyos 5Tree rings and arroyos 6Tree rings and arroyos 7Microplastic pollution discovered in St. Lawrence River sediments 2Scientists find how mysterious 'circular RNA' is formed, claim muscular dystrophy link 2
... tested two candidate vaccines that may eventually be used ... findings are published in the December 15 issue of ... Barney S. Graham, MD, PhD, and colleagues from the ... Maryland, the Fred Hutchinson Cancer Research Center in Seattle, ...
... the University of Rochester may have answered one of ... neurons, which are responsible for our crystal-clear thoughts, seem ... November issue of Nature Neuroscience, the Rochester study shows ... signals to represent the ambiguities of the real world--and ...
... may be fueling the spread of HIV in areas of ... two diseases, while HIV may be playing a role in ... according to a new study by researchers at Fred Hutchinson ... findings, published in the Dec. 8 issue of Science, found ...
Cached Biology News:Progress made in HIV vaccine development 2Mysterious 'neural noise' actually primes brain for peak performance 2Mysterious 'neural noise' actually primes brain for peak performance 3Malaria may fuel spread of HIV in sub-Saharan Africa 2
Anti-MMP-25 (Membrane-type MMP-6, Mt6MMP), hinge region; rabbit host...
Desmosomal Protein Immunogen: Full length, native protein purified from human epidermal keratins. Storage: -20 C, Avoid Freeze/Thaw Cycles...
Rabbit polyclonal to MAL ( Abpromise for all tested applications). entrezGeneID: 4118 SwissProtID: P21145...
Rabbit polyclonal to FLIP ( Abpromise for all tested applications). entrezGeneID: 8837 SwissProtID: O15519...
Biology Products: