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:4/17/2015)... 17, 2015 Reed Technology and ... regulatory data management, preparation, and submission to the U.S. ... of a new Lot Distribution Report (LDR) service for ... remain compliant with the FDA’s new rule on electronic ... to monitor the volume and timing of particular biologics ...
(Date:4/17/2015)... April 17, 2015 Global Stem ... GTCbio Stem Cell Summit 2015 April 27 - 29, ... latest developments in all areas of stem cell research ... stem cells and regenerative medicine. , The conference ... as well as translational research in stem cell therapies, ...
(Date:4/16/2015)... CAMBRIDGE, Mass. (PRWEB) April 16, 2015 ... of smart data solutions driven by Semantic Web ... provider of specialist information and services for the ... today announced that its joint solution ‘Smart Data ... named a finalist for the 2015 Bio-IT ‘Best ...
(Date:4/16/2015)... Regis Technologies looks forward to being ... Research (AACR) annual meeting. Regis will exhibit at this ... April 18, in Philadelphia and running through April 22, ... the oldest and largest organization focused on cancer research. ... partners with survivors to promote awareness and research funding. ...
Breaking Biology Technology:Reed Tech Announces FDA Lot Distribution Report (LDR) Service for Manufacturers of Biologic Products 2Reed Tech Announces FDA Lot Distribution Report (LDR) Service for Manufacturers of Biologic Products 3Global Stem Cells Group To Join GTCbio Stem Cell and Regenerative Medicine Summit in Boston, April 27-29, 2015 2Global Stem Cells Group To Join GTCbio Stem Cell and Regenerative Medicine Summit in Boston, April 27-29, 2015 3Cambridge Semantics and Informa Named Bio-IT Award Finalist for Smart Data Lake Solution for Clinical Trial and Competitive Intelligence 2Cambridge Semantics and Informa Named Bio-IT Award Finalist for Smart Data Lake Solution for Clinical Trial and Competitive Intelligence 3Regis Technologies Joins American Association for Cancer Research Annual Meeting 2
... Physicians and scientists from Reproductive Medicine Associates of ... their groundbreaking research study entitled Cleavage Stage ... Blastocyst Biopsy Does Not: A Novel Paired Analysis ... The study, led by Richard T. Scott, M.D. ...
... NEW YORK, Oct. 19, 2011 Progenitor Cell Therapy ... NBS ) ("NeoStem" or the "Company"), today ... the demands associated with its contract development and manufacturing ... add to the PCT client list.  NeoStem is an ...
...  Science Applications International Corporation (SAIC) (NYSE: SAI ... the National Institute of Allergy and Infectious Diseases (NIAID), ... provide preclinical services for the development of biopharmaceutical products ... at the end of fiscal year 2011.  The total ...
Cached Biology Technology:Revolutionary IVF Study May Change the Way Embryos are Biopsied for Genetic Disease 2Revolutionary IVF Study May Change the Way Embryos are Biopsied for Genetic Disease 3Progenitor Cell Therapy, a Wholly-Owned Subsidiary of NeoStem, Inc., Announces Additions to Management 2Progenitor Cell Therapy, a Wholly-Owned Subsidiary of NeoStem, Inc., Announces Additions to Management 3SAIC Awarded Contract by National Institute of Allergy and Infectious Diseases 2
(Date:4/20/2015)... The announcement comes as demand for fire ... Ireland,s foremost records management company, has ... an impressive track record of clients within the first 12 ... management sector in Dubai . As a ... employ a further eight staff members at its Irish headquarters. ...
(Date:4/14/2015)... April 14, 2015  HYPR Corp. today announced ... ® ) Alliance tm , an industry consortium ... commit to share technology and collaborate to deliver ... interoperable, more secure and private, and easier to ... verification that protects sensitive user information and eliminates ...
(Date:4/13/2015)... 2015 According to a ... Market Forecast & Opportunities, 2020", the global biometrics market ... 14% till 2020. The biometrics market is garnering ... and review of biometric management systems. The constant ... with greater efficiency, are resulting in increasing installation ...
Breaking Biology News(10 mins):Glenbeigh Records Management Wins Contract With Dubai Islamic Bank 2Glenbeigh Records Management Wins Contract With Dubai Islamic Bank 3HYPR Corp. Joins the FIDO Alliance 2HYPR Corp. Joins the FIDO Alliance 3Global Biometrics Market to Cross US$ 21 Billion by 2020: TechSci Research 2
... In the war against infectious disease, identifying the ... Wang and his colleagues from the Biodesign Institute at ... individual virus particles. Their research opens the door to ... may further the study of a broad range of ...
... University researchers have demonstrated a way to dramatically simplify ... New testing instrumentation developed by Antonia Garcia ... costly and produce results in less time. Current ... of working with blood, saliva, urine and other biological ...
... Ind. - A biosensor utilizing black platinum and carbon ... a better understanding of how the plant hormone auxin ... an associate professor of agricultural and biological engineering and ... movement of auxin along a plant,s root surface in ...
Cached Biology News:Visualizing viruses: new research pinpoints tiny invaders 2Visualizing viruses: new research pinpoints tiny invaders 3Health diagnosis made simpler 2Health diagnosis made simpler 3Health diagnosis made simpler 4Sensor important to understanding root, seedling development 2
... Topo I, Wild Type ... Tris-HCl, 1 mM DTT, 200 µM EDTA, 20% ... topoisomerase I (Topo I) is a monomeric protein ... gene. It catalyzes the relaxation of both positive ...
RNase-Free DNase 50 l...
Request Info...
... is a DNA-dependent RNA polymerase which is ... It efficiently synthesizes in vitro transcripts from ... from a T7 promoter. Transcripts can ... in vitro translation, substrates in RNA processing ...
Biology Products: