Navigation Links
Genotyping Single Nucleotide Polymorphisms with Molecular Beacons

.

Methods

Real-time monitoring of polymerase chain reaction: Molecular beacons were synthesized by Stratagene. A detailed protocol for molecular beacon design is available on Stratagenes web site at http://www.stratagene.com/index.asp?catID=17. The nucleotide sequence of the molecular beacons was as follows: SNP11A: 5-fam-gcgcagagacggAacttgctctgaaactgcgc-DABCYL-3; SNP11G: 5-fam-gcgcagagacggGacttgctctgaaactgcgc-DABCYL-3. Underlined regions identify the probe sequences, and an A or G capital letter in bold indicate the A/G polymorphism. Primers FP 5-aacttcacctccccacatca-3 and RP 5-atctggggcacagaagcatt-3 (Research Genetics) were selected from the region flanking the SNP11 in the BAC and generated a 144-bp-long amplicon. Human genomic DNAs were isolated from peripheral white blood cells, and extractions were carried out with the Super QUIK-GENE DNA isolation kit (Analytical Genetic Testing Center). Each 50-ml PCR reaction contained 100 ng of human genomic DNA, 500 nM molecular beacon A or G, 400 nM of each primer, 0.2 mM dATP, 0.2 mM dCTP, 0.2 mM dGTP, 0.4 mM dUTP, 3.5 mM MgCl2, 2.5 U of AmpliTaq Gold DNA polymerase (Applied Biosystems), and 1 U of uracil-N-glycosylase (UNG). Reactions were performed in triplicate in PE Applied Biosystems GeneAmp(r) 5700 Sequence Detection System. The PCR conditions were as follows: Cycling was preceded by 2 minutes at 50C; 10 minutes at 95C, followed by 40 cycles of 15 seconds; and 1 minute at 60C. In addition, a dissociation curve was generated after the last cycle of the PCR reaction. The thermal protocol for dissociation is defined as a hold at 95C for 15 seconds, a hold at 60C for 20 seconds, and a slow ramp (20 minutes) f
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
2. Purified Single-Stranded M13 DNA for Automated Sequencing
3. Detect Single Nucleotide Sequence Changes with Mx4000 Molecular Beacon Allelic Discrimination Kits
4. XL10-Gold Cells Supplied in New Single-Reaction Format
5. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
6. Single-cell RT-PCR Analysis of Paramecium Primaurelia with the Eppendorf cMaster RTplusPCR System
7. RNA, DNA, and Protein from a Single Sample
8. PFGE Analysis of Chromosomal DNA From a Single Hair Root
9. Single-Cell Complementation of Barley Mlo Mutants Using a PDS-1000/He Hepta System
10. Transient Gene Expression Following Single and Double Exponential Decay Electroporation Pulses
11. Simultaneous Detection of CYP3A4, CYP2D6 and CYP2C9 Metabolites with a Single, Sensitive, LC/MS/MS Method
Post Your Comments:
(Date:12/17/2014)... (PRWEB) December 17, 2014 Gene ... that it has entered into a technology access ... (ADM) to apply DNA2.0’s proprietary protein engineering technology, ... process. , “We are extremely excited that the ... platform. This proprietary bioengineering technology has now ...
(Date:12/17/2014)... SOUTH SAN FRANCISCO, Calif. , Dec. 17, ... positive results of a Phase 2 study evaluating ... for the treatment of patients with severe, chronic ... the current standard of care, including topical steroids ... endpoint was percent change in Visual Analog Scale ...
(Date:12/15/2014)...  GlassesOff Inc. (OTCBB: GLSO) announced today the appointment ... director of the Company,s Board of Directors. ... its CEO until its acquisition by Stanley Black ... Recognized as the inventor of the first Wi-Fi-based Active ... -based RFID solutions focused on improving operational efficiency, safety ...
(Date:12/13/2014)... (PRWEB) December 12, 2014 Clarassance, a ... announced its new name: Therabron Therapeutics , Inc. ... and bronchioles (a type of structure in the lungs ... company’s mission to develop novel protein therapeutics for the ... directors decided to change the name to mark the ...
Breaking Biology Technology:ADM and DNA2.0 Enter Into Protein Engineering Technology Access and Service Agreement 2Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 2Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 3Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 4AeroScout Founder Joins GlassesOff's Board of Directors 2AeroScout Founder Joins GlassesOff's Board of Directors 3Maryland-based Biotech Company's Path Forward in Treating Respiratory Diseases Sparks Name Change 2
... Varian, Inc.,(NasdaqGS: VARI) announced today the ... (ELSD) for conventional and high speed liquid,chromatography. ... solvent,compatibility, and excellent baseline stability, Varian ELSDs ... detectors for busy pharmaceutical,and analytical laboratories. In ...
... medium and ... large veins, ... ) today announced CE Mark approval of its ACUITY(R),Spiral left ventricular ... pacemakers, both of,which treat heart failure. The product features a spiral ...
... 3 NicOx S.A.,(Euronext Paris: COX) today announced ... the United States, which will assess the blood,pressure ... naproxen,using the Ambulatory Blood Pressure Monitoring (ABPM) technique. ... will together recruit a,total of around 420 osteoarthritis ...
Cached Biology Technology:New Chromatography Detectors From Varian, Inc. Extend the Range of Universal Detection for HPLC and LC/MS 2Boston Scientific Announces European Approval of New Heart Failure Lead 2Boston Scientific Announces European Approval of New Heart Failure Lead 3NicOx Initiates Two Large ABPM Studies for Naproxcinod in Hypertensive Patients with Osteoarthritis 2NicOx Initiates Two Large ABPM Studies for Naproxcinod in Hypertensive Patients with Osteoarthritis 3NicOx Initiates Two Large ABPM Studies for Naproxcinod in Hypertensive Patients with Osteoarthritis 4NicOx Initiates Two Large ABPM Studies for Naproxcinod in Hypertensive Patients with Osteoarthritis 5
(Date:12/10/2014)... Dec. 08, 2014 Research and Markets ... of the "Biometrics Market in Japan 2014-2018" ... http://photos.prnewswire.com/prnh/20130307/600769 The integration ... as rural banking and upgradation of the driver,s ... in the market. Besides the aforementioned projects, biometrics ...
(Date:12/5/2014)... -- Tute Genomics, a leader in cloud-based solutions for genome ... led by UK-based Eurovestech. Peak Ventures and a number ... "We are at a pivotal time in ... seeks new approaches for the diagnosis, treatment and prevention ... and CEO of Tute Genomics. "One of the main ...
(Date:12/3/2014)... , Dec. 2, 2014 As part ... readers, Inception Technologies is pleased to announce the release ... for customers to collect the workforce data that they ... that have been left by existing readers. Many such ... interface, connectivity and modern technology. Older models force users ...
Breaking Biology News(10 mins):Biometrics Market in Japan 2014-2018: Key Vendors are DDS, Fujitsu, Hitachi and NEC 2Tute Genomics Raises $2.3 Million to Scale Up Cloud-Based Genome Analytics and Accelerate Personalized Medicine 2Tute Genomics Raises $2.3 Million to Scale Up Cloud-Based Genome Analytics and Accelerate Personalized Medicine 3Inception Technologies to Release New Biometric Reader 2
... LANSING, Mich. -- The discovery of a new gene ... molecular factories of the future. A team of ... the discovery of Clumped Chloroplasts a new class ... Proceedings of the National Academy of Sciences . ...
... understand how a bacterial pathogen continues to damage tomatoes despite ... Pseudomonas syringae pv. tomato is the causative agent ... a disease that occurs worldwide and causes severe reduction in ... In the spring of 2010, for example, an ...
... plasticity and aging" team, directed by Jean-Marc Lemaitre, ... de Montpellier 1 and 2), has recently succeeded ... 100). These old cells were reprogrammed in vitro ... rejuvenated and human embryonic stem cells (hESC): cells ...
Cached Biology News:Discovery of new gene could improve efficiency of molecular factories 2Research team unravels tomato pathogen's tricks of the trade 2Research team unravels tomato pathogen's tricks of the trade 3Erasing the signs of aging in cells is now a reality 2Erasing the signs of aging in cells is now a reality 3
FluoSpheres® biotin-labeled microspheres, 1.0 µm, yellow-green fluorescent (505/515) *1% solids*...
chk 1 kinase...
muscle actin (HUC1-1)...
... The Human Neural Stem Cell ... media supplements, which can be used ... expansion of NSCs and the induction ... oligodendrocyte lineages. An antibody panel consisting ...
Biology Products: