Navigation Links
Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A

Specifically, quantitative real-time RT-PCR on the iCycler iQ was performed in duplicate or triplicate on 1 l of template cDNA per 20 l reaction. Mix A reactions consisted of PCR buffer (16.6 mM (NH4)2SO4, 67 mM Tris, pH 8.8, 6.7 mM MgCl2, 10 mM β-mercaptoethanol; Loging et al. 2000), 1 mM dNTPs (Invitrogen), 0.5 U of Platinum Taq DNA polymerase (Invitrogen), 10 nM fluorescein calibration dye (Bio-Rad), 1 l of a 1:1,500 dilution of 10,000x SYBR Green I stock, 500 nM of each GSP, and 1 l of cDNA. iQ supermix reactions consisted of iQ supermix (Bio-Rad) at a final concentration of 1x, 10 nM fluorescein calibration dye, 1 l of a 1:1,500 dilution of 10,000x SYBR Green I stock, 500 nM of each GSP, and 1 l of cDNA. To control for pipetting losses, 19 l of each 20 l reaction was amplified in a 96- well thin-wall PCR plate (Bio-Rad) using the following PCR parameters: 95C for 2 min followed by 50 cycles of 95C for 15 sec, 60C for 15 sec, and 72C for 15 sec. Melt-curve analysis was performed immediately following amplification by increasing the temperature in 0.4C increments starting at 65C for 85 cycles of 10 sec each. The presence of a single PCR product was verified both by the presence of a single melting temperature peak representing a specific product (vs. a nonspecific primer-dimer peak) using iCycler iQ analysis software and by detection of a single band of the expected size on a 4.5% Super AcrylAgarose gel.

Real-time RT-PCR was performed in duplicate or triplicate reactions. Each GSP pair was used with each reaction mix on each of the two different cDNA templates (derived from macula or periphery). Real-time RT-PCR reactions for detection of the endogenous control gene, A
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Rapid, High Throughput Mutation Discovery Using Fluorescent SSCP on the ABI PRISM 3100 and 3100Avant Genetic Analyzers
2. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
3. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
4. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
5. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
6. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
7. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
8. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
9. Real-Time Immuno-PCR on the iCycler iQ System, Rev A
10. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
11. Real-Time PCR: General Considerations, Rev A
Post Your Comments:
(Date:1/14/2014)... Canada (PRWEB) January 14, 2014 In ... its motives and methods in product development and promotion ... the industry. This mistrust, fueled by concerns about the ... fed by reports of spectacular fines to the world’s ...
(Date:1/14/2014)... , Jan. 14, 2014   Kinaxis ®, provider of ... ) and sales and operations planning ( S&OP ) service, ... Biomanufacturing Summit , which will be held at the Hilton ... conference, join Kinaxis customer Elisabeth Kaszas , Director of ...
(Date:1/14/2014)... 2014 During the 1600’s through the 1800’s ... Doctor’s Plague.” In this time period, doctors did not know ... times, to the death of vulnerable patients. In the same ... they may be unwittingly transmitting herpes viruses to their patients. ...
(Date:1/14/2014)... 2014 Holloway America today announced an ... test reports (MTRs) for all pressure vessels and tank ... is particularly important for equipment that must meet ASME, ... features search functionality that allows authorized users to quickly ...
Breaking Biology Technology:The Sunshine Act: Necessary Regulation or Unnecessary Dysregulation? New Life Science Webinar Hosted by Xtalks and IRB Services 2The Sunshine Act: Necessary Regulation or Unnecessary Dysregulation? New Life Science Webinar Hosted by Xtalks and IRB Services 3Event Alert: Kinaxis Customer to Present at the Biomanufacturing Summit "Supply Chain: Improving Network Effectiveness" 2Study: Fatigued Medical Interns Infect Their Patients with Herpes Viruses; The CBCD Sees a Parallel with “The Doctor’s Plague” 2Study: Fatigued Medical Interns Infect Their Patients with Herpes Viruses; The CBCD Sees a Parallel with “The Doctor’s Plague” 3Pressure Vessel Design and Fabrication Company Builds Database for Material Test Reports 2Pressure Vessel Design and Fabrication Company Builds Database for Material Test Reports 3
... JOLLA, Calif., Nov. 22, 2011  If algae is to be ... workers are needed to make that a reality. Applications are ... California San Diego to retrain workers as general science technicians ... and Imperial County region. Approximately 55 students ...
... Sequenom, Inc. (NASDAQ: SQNM ), a life ... Company,s participation at the 23rd Annual Piper Jaffray Healthcare Conference ... 2011. Ronald M. Lindsay, Ph.D., Director and ... will present on Tuesday, November 29, 2011 beginning at 8:30 ...
... Among the many hurdles to be cleared before human embryonic ... or not transplanted cells can functionally integrate into target organs ... Proceedings of the National Academy of Sciences, a ... lab from blank slate human embryonic stem cells and implanted ...
Cached Biology Technology:Free Job Training: 55 Grants Available to Retrain Workers for Biofuels Industry 2Free Job Training: 55 Grants Available to Retrain Workers for Biofuels Industry 3Sequenom Announces Participation at the 23rd Annual Piper Jaffray Healthcare Conference 2Implanted neurons, grown in the lab, take charge of brain circuitry 2Implanted neurons, grown in the lab, take charge of brain circuitry 3
(Date:4/17/2014)... trials to treat tuberculosis could be the basis for ... bacteria, fungal infections and parasites, yet evade resistance, according ... collaborators. , Led by U. of I. chemistry professor ... drug SQ109 attacks the tuberculosis bacterium, how the drug ... to malaria and how targeting multiple pathways reduces ...
(Date:4/17/2014)... modeling study suggests that fish consumption advisories for ... to long-lived contaminants like persistent organic pollutants (POPs). ... of researchers including University of Toronto Scarborough PhD ... at how different levels of environmental contamination, a ... chemicals in the body influenced exposure in her ...
(Date:4/17/2014)... University engineer has developed a patented technique that ... devices. The same technique could help police during ... and Kay L. Theede chair in engineering and ... his research team have created a template-based system ... trunks. The distance detection method called stand-off ...
Breaking Biology News(10 mins):Multitarget TB drug could treat other diseases, evade resistance 2Fish consumption advisories fail to cover all types of contaminants 2Patented research remotely detects nitrogen-rich explosives 2
... Sharing is a behavior on which day care workers and kindergarten ... for our ape cousins the bonobos, sharing just comes naturally. ... Current Biology, it looks like bonobos never seem to ... notorious for hogging food to themselves, by physical aggression if necessary. ...
... Imagine that you are emerging from the subway and heading ... in the wrong direction. For a moment, you feel disoriented, ... surrounding streets quickly helps you pinpoint your location, and you ... Research tells us that human adults, toddlers, rats, chicks ...
... the first to design a bioactive nanomaterial that promotes the ... of expensive growth factors. Minimally invasive, the therapy activates the ... therapy can do this. The results will be published ... of the National Academy of Sciences (PNAS). "Unlike ...
Cached Biology News:'Peter Pan' apes never seem to learn selfishness 2'Peter Pan' apes never seem to learn selfishness 3Ability to navigate may be linked to genes, researcher says 2Ability to navigate may be linked to genes, researcher says 3Growing cartilage -- no easy task 2
Hybridization Buffer 3.6 ml...
Chicken polyclonal to Thaumatin 2 ( Abpromise for all tested applications). Antigen: Full length Thaumatin 2...
Proteinase K Buffer 3.9 ml...
Rabbit polyclonal to VEGF Receptor 2 (phospho Y1214) ( Abpromise for all tested applications). entrezGeneID: 3791 SwissProtID: P35968...
Biology Products: