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:12/17/2014)... 2014 “Never say, ‘I cannot ... Marburg’s Department of Physics, advises young women pursuing ... released SPIE Women in Optics 18-month planner ... to ask questions.” , In the annual planner, ... occupations ranging from university professor and laboratory researcher ...
(Date:12/17/2014)... This report is a professional ... For an overview analysis, the report introduces quartz ... structure, industry overview, policy analysis, and news analysis, ... has witnessed rapid development with key manufacturers increasing ... report mentions quartz tubing upstream raw materials and ...
(Date:12/17/2014)... -- Vermillion, Inc. (NASDAQ: VRML ), a bio-analytic ... next step in its expanded strategy to rebuild the ... naming of current Chief Operating Officer, Valerie Palmieri ... Chairman of the Board and CEO James LaFrance ... are effective January 1, 2015. "Jim and ...
(Date:12/17/2014)... --  Synageva BioPharma Corp. (NASDAQ: GEVA ... disorders, announced today its presentation at the upcoming J.P. ... Francisco, CA. Sanj K. ... on Monday, January 12, 2015, at 9:00 a.m. PST ... and may be accessed from the "Webcasts & Presentations" ...
Breaking Biology Technology:Network, Multitask, Persevere: SPIE Women in Optics Planner Subjects Share from Experience 2Network, Multitask, Persevere: SPIE Women in Optics Planner Subjects Share from Experience 32020 Quartz Tubing Market: Global Development & Forecasts Analysis Now at DeepResearchReports.com 2Vermillion Names Valerie Palmieri CEO; James LaFrance to continue as Chairman of Board 2Vermillion Names Valerie Palmieri CEO; James LaFrance to continue as Chairman of Board 3Vermillion Names Valerie Palmieri CEO; James LaFrance to continue as Chairman of Board 4
... MOUNTAIN VIEW, Calif., Dec. 4 Based on ... & Sullivan recognizes Honeywell Enraf with the 2008 ... Honeywell Enraf,s comprehensive suite of blending products facilitates ... and other materials, but also minimizes the end-products, ...
... of Life Technologies Corporation (NASDAQ:LIFE), a provider of ... announced that its technology for next-generation genomic analysis, ... Science Innovation of the Year for 2008 by ... the life science industry. In making the announcement ...
... an estimated ten percent of pregnant women in the U.S. ... cigarette byproducts from mothers who smoke affects an estimated 420,000 ... , Now, in the first study of its kind, a ... umbilical cord blood to better understand the fetal health risks ...
Cached Biology Technology:Frost & Sullivan Commends Honeywell Enraf for Cutting-Edge Products and Post-Sales Support 2Frost & Sullivan Commends Honeywell Enraf for Cutting-Edge Products and Post-Sales Support 3 Applied Biosystems SOLiD System Named Life Science Innovation of the Year : The Scientist Magazine Cites the Technology's High Throughput, Lower Sequencing Cost, and Rapid Technology Innovation for Unlocking New Approaches to Clinical Research 2 Applied Biosystems SOLiD System Named Life Science Innovation of the Year : The Scientist Magazine Cites the Technology's High Throughput, Lower Sequencing Cost, and Rapid Technology Innovation for Unlocking New Approaches to Clinical Research 3 Applied Biosystems SOLiD System Named Life Science Innovation of the Year : The Scientist Magazine Cites the Technology's High Throughput, Lower Sequencing Cost, and Rapid Technology Innovation for Unlocking New Approaches to Clinical Research 4 Applied Biosystems SOLiD System Named Life Science Innovation of the Year : The Scientist Magazine Cites the Technology's High Throughput, Lower Sequencing Cost, and Rapid Technology Innovation for Unlocking New Approaches to Clinical Research 5Cutting the cord to determine babies' health risk from toxic exposure 2Cutting the cord to determine babies' health risk from toxic exposure 3Cutting the cord to determine babies' health risk from toxic exposure 4
(Date:12/10/2014)... , December 9, 2014 ... ITN,s BCARD Platform   ... enterprise meeting scheduling solutions for business-to-business (B2B) events, ... world leader in mobile near-field communication (NFC), Bluetooth ...      (Logo: http://photos.prnewswire.com/prnh/20140612/691055) , Jifflenow ...
(Date:12/3/2014)... IRVINE, Calif. , Dec. 2, 2014 /PRNewswire/ ... of globally deployed, innovative test solutions for military, ... the newest version of its successful TS-900 ... the performance and features of high-end systems to ... with outstanding value compared to traditional ATE. ...
(Date:11/21/2014)... , Nov. 20, 2014 Strict laws against ... are piloting the North American and European automotive sector ... growing, gesture recognition systems that are intuitive and able ... in the industry. New analysis from ... Recognition Market in Europe and ...
Breaking Biology News(10 mins):Jifflenow And ITN International Bring Cutting-Edge Badge Scanning Technology To B2B Events 2Jifflenow And ITN International Bring Cutting-Edge Badge Scanning Technology To B2B Events 3Marvin Test Solutions Brings New Capabilities to PXI-Based Semiconductor Test with TS-960 2Marvin Test Solutions Brings New Capabilities to PXI-Based Semiconductor Test with TS-960 3Gesture Recognition Intensifies with the Rise of In-car Smartphone Integration, Says Frost & Sullivan 2Gesture Recognition Intensifies with the Rise of In-car Smartphone Integration, Says Frost & Sullivan 3Gesture Recognition Intensifies with the Rise of In-car Smartphone Integration, Says Frost & Sullivan 4
... can often only be solved using forensics to piece ... Helmut Schmitz from the Institute of Zoology at the ... Peter Grnberg Institut at the Forschungzentrum Jlich went through ... beetles of the genus Melanophila and their ...
... This press release is available in ... I,m depressed and I,m depressed because I eat" in the ... research being undertaken by scientists affiliated with the University of ... Medicine. "In addition to causing obesity, rich foods can actually ...
... the selective force that preserves adaptive traits in organisms and ... well known that evolutionary change in organisms also trigger fundamental ... prime example of this evolutionary feedback loop in a few ... Colonial times trapped a fish called the alewife. ...
Cached Biology News:Forensics ferret out fire beetle secret 2Forensics ferret out fire beetle secret 3Scientists start explaining Fat Bastard's vicious cycle 2Not a 1-way street: Evolution shapes environment of Connecticut lakes 2
Rat Epidermal Keratinocytes (REK) (>500,000 cells)...
anti-Yan 8B12H9...
sPLA2-XII Human, Rabbit Polyconal Antibody Anti-Human sPLA2-XII Antibody was raised in rabbits against Human sPLA2-XII His-Tagged Fusion Protein Antigen: The recombinant human protein sPLA-XII...
... Expand the computing power of your ValveBank ... sequences with a PC computer running Windows 98, ... load unlimited programs to disk, print out program ... memory in one simple step., EasyCode is used ...
Biology Products: