Navigation Links
Cells-to-cDNA II Using FACS Sorted Cells

Fluorescence activated cell sorting (FACS) is a tool for measuring phenotypic, biochemical and molecular properties of individual cells using fluorescent probes that bind specifically to molecules associated with the cell. This technology is commonly used to collect cells of a given population down to the level of a single cell.

Figure 1.Linear Amplification from 1-1000 Cells.

Dr. Daniel Douek of the NIH Vaccine Research Center recently used Ambion's Cells-to-cDNA II Lysis Buffer to analyze gene expression of human -glucuronidase (hGUS) in FACS sorted Jurkat cells using real-time RT-PCR. GUS is an enzyme constituitively expressed at moderate levels in all cells.

Jurkat cells were sorted by FACS and collected at 1, 10, 100 and 1000 cells per well in a 96-well PCR plate. Each well was preloaded with 4 l of Cells-to-cDNA II Lysis Buffer. After the cells were sorted and lysed, the plate was placed at -80C for 1 week. The lysates were then DNase treated according to the Cells-to-cDNA II protocol. One-step RT-PCR was performed in 20 l using a TaqMan probe for hGUS. Six replicates were run for each cell concentration. A representative amplification plot is shown in Figure 1. Results show quantitative amplification from 1-1000 cells, demonstrating a 3-log dynamic range and reproducibility of the assay.

TaqMan is a registered trademark of Applied Biosystems.

Ordering Information
Cat# Product Name Size 1722 Cells-to-cDNA II Kit 40 rxns 1723 Cells-to-cDNA II Kit 100 rxns 8723 Cells-to-cDNA II Cell Lysis Buffer 5 x 10 ml

Related Articles

Go From Cells to cDNA Directly: Using FACS Samples
Real-time PCR Goes Prime Time
Real-Time Relative RT-PCR: How It's Done
Synthesize cDNA Directly from Limiting Samples
Quick Real-time Analysis of Small Cell Samples



Page: All 1 2

Related biology technology :

1. Cells-to-cDNA II Applications
2. Your Data: Detection of Oocyte-specific Gene Expression Using Ambions Cells-to-cDNA Kit
3. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
4. Oil-Free PCR Using the Hot Top Assembly
5. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
6. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
7. Using XL1-Red Mutator Strain to Generate Esterase Variants
8. Detect Released Oligosaccharides Using the Eagle Eye II Still Video Imaging System
9. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
10. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
11. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
Post Your Comments:

(Date:6/24/2016)... NC (PRWEB) , ... June 24, 2016 , ... Researchers ... the most commonly-identified miRNAs in people with peritoneal or pleural mesothelioma. Their findings are ... to read it now. , Diagnostic biomarkers are signposts in the blood, lung ...
(Date:6/23/2016)...   Boston Biomedical , an industry leader ... target cancer stemness pathways, announced that its lead ... Designation from the U.S. Food and Drug Administration ... gastroesophageal junction (GEJ) cancer. Napabucasin is an orally ... stemness pathways by targeting STAT3, and is currently ...
(Date:6/23/2016)... 23, 2016 Houston Methodist Willowbrook Hospital ... Sports Association to serve as their official health ... Methodist Willowbrook will provide sponsorship support, athletic training ... association coaches, volunteers, athletes and families. ... Sports Association and to bring Houston Methodist quality ...
(Date:6/23/2016)... 2016  The Biodesign Challenge (BDC), a university competition ... harness living systems and biotechnology, announced its winning teams ... New York City . The ... projects at MoMA,s Celeste Bartos Theater during the daylong ... senior curator of architecture and design, and Suzanne ...
Breaking Biology Technology:
(Date:4/26/2016)... LONDON , April 26, 2016 /PRNewswire/ ... Systems, a product subsidiary of Infosys (NYSE: ... partnership to integrate the Onegini mobile security platform ... ) The integration ... security to access and transact across channels. Using ...
(Date:4/14/2016)... Israel , April 14, 2016 ... Authentication and Malware Detection, today announced the appointment of ... assumed the new role. Goldwerger,s leadership appointment ... on the heels of the deployment of its platform ... BioCatch,s behavioral biometric technology, which discerns unique cognitive and ...
(Date:3/23/2016)... 23, 2016 ... Gesichts- und Stimmerkennung mit Passwörtern     ... MESG ), ein führender Anbieter digitaler ... mit SpeechPro zusammenarbeitet, um erstmals dessen Biometrietechnologie ... die Möglichkeit angeboten, im Rahmen mobiler Apps ...
Breaking Biology News(10 mins):