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:9/16/2014)... Pharma Packaging Solutions announced ... for their 89th year in business. Adding ... Packaging Solutions is preparing packaging operations changes ... new procedures. Establishing training and system validation ... Pharma Packaging Solutions will be ready to ...
(Date:9/16/2014)... NEW ORLEANS , Sept. 16, 2014 ... Austin, Texas -based cybersecurity company focused on ... 2FA ONE, will support the ability to generate a ... As a pioneer of user-based RFID authentication, ... used for physical access, for many years. RFID authentication ...
(Date:9/15/2014)... , Sept. 15, 2014   Global ... innovator of scientific products and services for satellite ... Acquisition Solution for Integrated Services (OASIS) from the ... GST was selected to provide the government best-value ... Small Business (SB) category in Pool 4. The ...
(Date:9/15/2014)... go downhill fast when a patient has sepsis, a ... a patient,s blood -- often too fast for antibiotics ... spleen and developed by a team at Harvard,s Wyss ... way doctors treat sepsis. , "Even with the ... care units at least 30 percent of the time," ...
Breaking Biology Technology:Adding Serialization Capabilities and Expanded Capacity, Pharma Packaging Solutions Is Preparing for 89th Year of Business 22FA, Inc. Announces RFID-based One-Time Password (OTP) Authentication 2Global Science & Technology, Inc. Awarded GSA OASIS Small Business Contract 2Global Science & Technology, Inc. Awarded GSA OASIS Small Business Contract 3Blood-cleansing biospleen device developed for sepsis therapy 2Blood-cleansing biospleen device developed for sepsis therapy 3Blood-cleansing biospleen device developed for sepsis therapy 4
... April 27 BioRN Cluster Management GmbH presents ... management,s collaborative project, a web-based software to manage complex ... or single companies with an easy-to-use application for the ... the core needs of the different project partners: of ...
... , April 27 A non-profit organization is urging PepsiCo ... amount of its corporate donations.  "Unlike Coca-Cola, Pepsi refuses to divulge the ... Quinlan , President of Parents and Friends of Ex-Gays & Gays (PFOX). ... "This secrecy by ...
... , ... partners with RURO to offer GLP/GMP certified hosting services. , ... Annapolis, MD (Vocus) April 27, 2010 -- ... with laboratory software provider RURO. RURO’s leading edge laboratory applications can now be ...
Cached Biology Technology:Web-Based Management Suite Supports Complex Research Consortia 2Pepsi Urged to Follow Example of Coca-Cola and Stop Hiding Corporate Donations 2Sidus BioData Partners with Life Science Software Provider RuRo 2
(Date:9/15/2014)... epilepsy drug to a morphine regimen can result in ... combination can reduce the dosage of the opioid needed ... researchers at Indiana University. , The result could bring ... difficult-to-treat condition often felt in the arms and legs ... a huge unmet need for better treatments for neuropathic ...
(Date:9/15/2014)... for both plants and animals alike. , Understanding just ... lead to the design of better drugs for diabetes ... and vegetables farmers are able to grow. Stanford University ... the cell by piecing together proteins slightly wider than ... To determine the size, shape and orientation of one ...
(Date:9/15/2014)... When this week,s print issue of the journal ... up from New Mexico, Montana and even the Netherlands, ... more and more the norm in these connected times. ... it will produce innovations in biology, medicine, biotechnology and ... this scientist talked with that one, that one knew ...
Breaking Biology News(10 mins):IU study: Combining epilepsy drug, morphine can result in less pain, lower opioid doses 2X-rays unlock a protein's SWEET side 2X-rays unlock a protein's SWEET side 3Collaboration drives achievement in protein structure research 2
... named beneficiaries of the Joshua E. Neimark Memorial Travel ... young children,s interest in insects, an effort to fine-tune ... a way to lower the cost of a key ... financial support to attend America,s largest general scientific conference, ...
... Biologists at UC San Diego have identified the specific region ... development. Their discovery, which appears in a paper in ... is a critical first step for the development of ... leukemia, multiple myeloma, anemia and a host of other diseases ...
... the California Institute of Technology (Caltech) have obtained the ... fruit fly. The workby Michael Dickinson, the ... with postdoctoral scholars Gaby Maimon and Andrew Strawsuggests that ... fly (Drosophila melanogaster) "is in a different and more ...
Cached Biology News:Joshua E. Neimark Memorial Travel Assistance Award winners 2Biologists image birth of blood-forming stem cells in embryo 2Biologists image birth of blood-forming stem cells in embryo 3Caltech researchers obtain first brain recordings from behaving fruit flies 2Caltech researchers obtain first brain recordings from behaving fruit flies 3
... optically clear polystyrene with frosted labeling ... for cell counting. 100% integrity tested. ... new product number, created to easily ... no availability yet, please order under ...
Packaging: package with 500 l Patent/trademark/license (electronic): (r) Registered trademark of Molecular Probes...
... VersaDoc MP 4000 System is a ... system capable of supporting a wide ... chemiluminescence, densitometry, and chemiflourescence. Red, ... imaging of multiplexed fluorescent samples, reducing ...
... hydrophobic barrier pen for immunohistochemistry and in ... frozen or paraffin-embedded tissue sections mounted on ... heat-stable, water-repellent barrier that keeps staining reagents ... of reagents when differentially staining two sections ...
Biology Products: