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


'"/>

Source:


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:
*Name:
*Comment:
*Email:


(Date:10/9/2017)... ... October 09, 2017 , ... At its national board ... Stubbs, a professor in Harvard University’s Departments of Physics and Astronomy, has been selected ... member of the winning team for the 2015 Breakthrough Prize in Fundamental physics for ...
(Date:10/9/2017)... ... October 09, 2017 , ... ... medical journal, Epilepsia, Brain Sentinel’s SPEAC® System which uses the surface electromyography ... generalized tonic-clonic seizures (GTCS) using surface electromyography (sEMG). The prospective multicenter phase ...
(Date:10/7/2017)... ... October 06, 2017 , ... ... launched its ProxiMeta™ Hi-C metagenome deconvolution product, featuring the first commercially available ... bioinformatics software to perform Hi-C metagenome deconvolution using their own facilities, supplementing ...
(Date:10/6/2017)... Mass. (PRWEB) , ... October 06, 2017 , ... ... female entrepreneurship within the healthcare and technology sector at their fourth annual Conference ... panels featuring 30 inspiring speakers and the ELEVATE pitch competition showcasing early stage ...
Breaking Biology Technology:
(Date:5/16/2017)... DALLAS , May 16, 2017   ... for health organizations, and MD EMR Systems ... certified development partner for GE, have established a ... Patient Portal product and the GE Centricity™ products, ... Centricity EMR. These new integrations ...
(Date:4/24/2017)... Janice Kephart , former 9/11 ... Partners, LLP (IdSP) , today issues the following ... March 6, 2017 Executive Order: Protecting the ... be instilled with greater confidence, enabling the reactivation ... applications are suspended by until at least July ...
(Date:4/18/2017)... 18, 2017  Socionext Inc., a global expert in SoC-based imaging ... server, the M820, which features the company,s hybrid codec technology. A ... Tera Probe, Inc., will be showcased during the upcoming Medtec Japan ... at the Las Vegas Convention Center April ... Click here for ...
Breaking Biology News(10 mins):