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:9/18/2014)... -- Research and Markets has announced the ... 2014" report to their offering. ... professional and in-depth study on the current state of ... provides a basic overview of the industry including definitions, ... market analysis is provided for the international markets including ...
(Date:9/18/2014)... , Sept. 18, 2014  U.S. biotech company ... an exclusive license for technology developed by the ... expression in plants, including for applications in production of ... The technology was developed under the direction of ... Kultevat and former president of the Danforth Plant Science ...
(Date:9/18/2014)... , Sept. 18, 2014 /PRNewswire/ - Oncothyreon Inc. (NASDAQ: ... previously announced concurrent but separate underwritten offerings of 10,000,000 ... the public of $2.00 per share, for expected gross ... Series A Convertible Preferred Stock at a price to ... proceeds of $20 million. Each share of Series A ...
(Date:9/18/2014)... (PRWEB) September 18, 2014 OMICS ... the globe to participate in the ‘Open Access ... articles to OMICS Group Pharma Journals ... one of the fastest emerging Sciences, accelerating with ... healthcare services across the globe. The industry is ...
Breaking Biology Technology:Global Human Albumin Industry Report 2014 2Global Human Albumin Industry Report 2014 3Kultevat obtains license of gene switch technology 2Oncothyreon Prices $40 Million Concurrent Public Offerings of Common Stock and Preferred Stock 2Oncothyreon Prices $40 Million Concurrent Public Offerings of Common Stock and Preferred Stock 3Pharma and Omics Journals to Address Global Challenges during Open Access Week 2Pharma and Omics Journals to Address Global Challenges during Open Access Week 3
... to Remain as Advisor to Company, FRAZER, Pa., ... its Executive Vice President, General Counsel &,Secretary, John E. ... effective March 31. Mr. Osborn has agreed to remain ... policy matters., "John has been an invaluable member ...
... Inc. (Nasdaq:,EVVV) will release its 2007 fourth quarter and ... market trading hours., The company will host a ... CT (8:30 a.m. ET). ev3 management will review its ... outlook, followed,by a question and answer session., The ...
... Spanish . , ANN ARBOR, Mich.---A new ... to operate a portable GPS locator, a cell phone, ... involving the University of Michigan shows. , A report ... issue of the journal Science. Authors include researchers from ...
Cached Biology Technology:Cephalon General Counsel John E. Osborn to Resign Position 2Cephalon General Counsel John E. Osborn to Resign Position 3ev3 Inc. to Report 2007 Fourth Quarter and Full-Year Financial Results and Host Conference Call on February 22, 2008 2Knee brace generates electricity from walking 2
(Date:9/18/2014)... protective tentacles of host anemones, but new research shows ... across the open ocean. Although the process of long-distance ... the first time that the high level exchange of ... Steve Simpson, Senior Lecturer in Marine Biology and Global ... the Australian Research Council Centre of Excellence for Coral ...
(Date:9/17/2014)... UCLA atmospheric scientists were instrumental in the creation of ... tool to classify the fire threat potential of the ... a spark into an inferno. The index was introduced ... with UCLA and San Diego Gas and Electric. , ... to "extreme" that will be used to help fire ...
(Date:9/17/2014)... and his doctoral student have designed a device based ... from fog and dew., The device could provide water ... the globe., Cheng Luo, professor in the Mechanical & ... the same College of Engineering department, published "Bioinspired Plate-Based ... (American Chemical Society) Applied Materials & Interfaces ...
Breaking Biology News(10 mins):Expedition finds Nemo can travel great distances to connect populations 2Expedition finds Nemo can travel great distances to connect populations 3UCLA scientists play key role in developing new Santa Ana Wildfire Threat Index 2Shorebird's beak inspires UT Arlington research on water collection 2
... crystals, known to be a catalyst for heart attacks and ... can lead to the inflammation and hardening of arteries, according ... by George Abela, chief of the cardiology division in MSU,s ... new insights into how arteries harden a process called ...
... patches of woods in urban areas seem to provide adequate ... they fly between wintering and breeding grounds, new research has ... of cities worldwide, migrating landbirds increasingly must pass through vast ... on which many species rely. "The good news is ...
... N.Y. (May 18, 2010) -- Symmetry breaking events ... They are required for cell division, development, and movement ... Moreover, in multicellular organisms, symmetry breaking allows the generation ... arrangement of tissues and organs achieved during embryogenesis. ...
Cached Biology News:Cholesterol crystals incite inflammation in coronary arteries 2Even small patches of urban woods are valuable for migrating birds 2Even small patches of urban woods are valuable for migrating birds 3
... Pierce offers maleimide activated carrier proteins ... conjugates. Imject Maleimide Activated Carrier Proteins ... been preactivated with a heterobifunctional cross-linker ... stable, sulfhydryl-reactive carrier protein. Thus ...
...
... A perfect complement to high-speed analytical techniques, the AutoTrace SPE is a ... automates the cartridge conditioning, sample loading and elution steps for large volumes aqueous ... ... ...
Cell Culture Flask, 25 cm, tissue-culture treated polystyrene...
Biology Products: