Navigation Links
Real-Time Immuno-PCR on the iCycler iQ System, Rev A

rimer (5'-ACATATCCAGCCATGCACAC- 3'), and was purified with the QIAquick PCR purification kit (QIAGEN).

Real-time PCR was performed in the Bio-Rad iCycler iQ system. The primers used were 5'-GTTAGCCGGGCTGCACTC- 3' and 5'-ACATATCCAGCCATGCACAC-3', which produced a 71 bp product. Reaction volumes were 25 l, containing 1x PCR buffer (Sigma), 4 mM MgCl2 (Sigma), 200 M dNTPs (Sigma), 0.08 g/l BSA (Fermentas), 1 U JumpStart Taq polymerase (Sigma), 300 nM of each primer, and 0.5x SYBR Green I (Molecular Probes, Inc.) Cycling conditions were 95C for 3 min, followed by 40 cycles of 95C for 20 sec, 60C for 20 sec, and 72C for 25 sec. The fluorescence data used for quantitation were collected at the end of each 72C step.


Results and Discussion
The sandwich real-time immuno-PCR system was assembled in a standard PCR plate from Bio-Rad as shown in Figure 1. To each well, 50 l anti-PSA10 (10 g/ml in 0.2 M phosphate buffer) was added and allowed to adsorb to the surface of the PCR plate overnight. The wells were then washed three times with wash buffer 1 (0.154 M NaCl, 5 mM Tris, pH 7.75, 0.005% Tween 20, 0.1% Germall II), and the surface was then blocked from further adsorption by incubating at room temperature overnight with blocking buffer (50 mM Tris, pH 7.0, 6% D-sorbitiol, 0.1% BSA, 0.05 % NaN3). The wells were washed with wash buffer 1, followed by incubation at room temperature with 5 l of PSA standards (containing 2.4 x 106 to 2.8 x 109 molecules) and 20 l Tris-HCl buffered salt solution containing BSA for 1 hr (CanAg PSA EIA instructions). The samples were then washed three times with wash buffer 1. Biotinylated anti-PSA66 was diluted to 0.92
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. 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
2. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
3. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
4. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
5. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
6. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
7. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
8. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
9. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
10. Real-Time PCR: General Considerations, Rev A
11. Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System, Rev A
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Real Time Immuno PCR the iCycler System Rev

(Date:11/21/2014)... November 20, 2014 Offering a complete ... Sartorius Entris Balance promotion . The Sartorius Entris Balance ... durable, high accuracy, and easy-to-maintain balance entry level balance. ... It was designed to help customer’s bridge the gap ... balance that is very durable. Sartorius is the second ...
(Date:11/21/2014)... Palo Alto, Calif. (PRWEB) November 20, 2014 ... (CHI), a nonprofit, public policy research organization representing ... and pharmaceutical firms, and PwC US ... other states in life sciences industry growth. The ... and 2013. The 2015 California Biomedical Industry ...
(Date:11/21/2014)... The November 2014 issue of BioTechniques ... access journal featuring the following articles:, , ... photoresponsiveness in the retina , DNA from ... using chromogenic substrates in zebrafish , Re-use ... electrophoresis , Nuclear LC3-positive puncta in stressed ...
(Date:11/21/2014)... Los Angeles, CA (PRWEB) November 21, 2014 ... is it that makes Albert Einstein so instantly recognizable? ... South African-born physicist, mathematician, astronomer and author Hilton Ratcliffe ... that they have nothing at all to do with ... ( Muse Harbor Publishing , November 21, 2014), Ratcliffe ...
Breaking Biology Technology:Pipette.com Announces an Extension on the Sartorius Entris Balance Promotion to the End of 2014 2California Healthcare Institute and PwC Report Boom in California’s Biomedical Industry 2California Healthcare Institute and PwC Report Boom in California’s Biomedical Industry 3California Healthcare Institute and PwC Report Boom in California’s Biomedical Industry 4BioTechniques: The International Journal of Life Science Methods Announces the Publication of the November 2014 Issue—Available in Print and Online 2BioTechniques: The International Journal of Life Science Methods Announces the Publication of the November 2014 Issue—Available in Print and Online 3“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 2“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 3“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 4
... - A new technique inspired by elegant pop-up books ... to be mass-produced by the sheet. Devised by ... enables the rapid fabrication of not just microrobots, but ... 18 layers of carbon fiber, Kapton (a plastic film), ...
... Healthcare organizations are becoming more efficient and effective ... more connected patient care environment. But to have this ... the electronic medical record (EMR) and enterprise content management ... leader in ECM for healthcare , has a ...
... A protein that has shown early promise in preventing the ... has been found in a new study to be a key ... protein biglycan needs to be present to stabilize synapses at the ... Brown University that appears in the Feb. 14, 2012, issue of ...
Cached Biology Technology:In new mass-production technique, robotic insects spring to life 2In new mass-production technique, robotic insects spring to life 3Johns Hopkins Health System is 100th healthcare Provider to Integrate OnBase With Leading Electronic Medical Record System 2Johns Hopkins Health System is 100th healthcare Provider to Integrate OnBase With Leading Electronic Medical Record System 3When nerve meets muscle, biglycan seals the deal 2
(Date:11/7/2014)... -- Leading Biometric companies aim to provide consumers ... transactions.  Companies in focus today are: NXT-ID, Inc. (NASDAQ: ... ), Google Inc. (NASDAQ: GOOG ), Microsoft Corporation ... EBAY ) and MasterCard Inc. (NYSE: MA) NXT-ID, ... authentication company focused on the growing mobile commerce is pleased ...
(Date:11/5/2014)... Santamaria, associate professor of biology in the UTSA ... in the nation selected to receive a two-year ... for Exploratory Research (EAGER). The funding supports President ... researchers to create new technology that will demystify ... complex behaviors in neuroscience are broken into the ...
(Date:11/4/2014)... School of Medicine announced today that it is ... by the Bill & Melinda Gates Foundation. ... and Reproductive Sciences; and Vice Chair of Research ... Medicine will pursue an innovative global health and ... Assess Fetal Neurodevelopment.", Grand Challenges Explorations (GCE) funds ...
Breaking Biology News(10 mins):Universal Authentication Methods Provide More Secure Operations for Biometrics Devices - Company Joins FIDO Alliance for Consumer Protection 2Universal Authentication Methods Provide More Secure Operations for Biometrics Devices - Company Joins FIDO Alliance for Consumer Protection 3Universal Authentication Methods Provide More Secure Operations for Biometrics Devices - Company Joins FIDO Alliance for Consumer Protection 4Universal Authentication Methods Provide More Secure Operations for Biometrics Devices - Company Joins FIDO Alliance for Consumer Protection 5Universal Authentication Methods Provide More Secure Operations for Biometrics Devices - Company Joins FIDO Alliance for Consumer Protection 6UTSA biology professor awarded $300,000 NSF grant for brain research 2UTSA biology professor awarded $300,000 NSF grant for brain research 3Temple University School of Medicine receives Grand Challenges Explorations grant 2
... hiatus North America,s tallest bird, the statuesque whooping crane ( ... Bird House at the Smithsonian,s National Zoological Park. An 11-year-old ... in Florida and is now on exhibit in the nation,s ... native to the United States. There are only eight other ...
... form of vitamin E called alpha-tocotrienol can trigger production of ... cells, preventing those cells from dying after a stroke, new ... identified so far that this form of vitamin E uses ... natural substance might be more potent than drugs targeting single ...
... - One of the peskiest household pests, while disastrous to ... to a Purdue University study. Mike Scharf, the O. ... his laboratory has discovered a cocktail of enzymes from the ... the barriers that inhibit fuel production from woody biomass. The ...
Cached Biology News:National Zoo Welcomes Whooping Crane 2'Gifted' natural vitamin E tocotrienol protects brain against stroke in 3 ways 2'Gifted' natural vitamin E tocotrienol protects brain against stroke in 3 ways 3'Gifted' natural vitamin E tocotrienol protects brain against stroke in 3 ways 4Termites' digestive system could act as biofuel refinery 2
... the mycobacteria found in Complete Freund's Adjuvant so ... Freund's Adjuvant is used for ... to produce water-in-oil emulsions of immunogens. ... antibody responses which can be attributed to the ...
2,4-Dinitrophenyl hapten is conjugated to Keyhole Limpet (KLH) protein through lysine....
PGF1α-d4 contains 4 deuterium atoms at the 3, 3', 4, and 4' positions. It is intended for use as an internal standard for the quantification of PGF1α by GC- or LC-mass spectrometry....
...
Biology Products: