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:12/2/2016)... ... December 02, 2016 , ... Robots will storm the Prudential Center in ... 2016. The event, which is held on the United Nations International Day of Persons ... back into the workplace. Suitable Technologies is partnering with NTI to showcase how technology ...
(Date:11/30/2016)... , ... November 30, 2016 , ... ... moving magnet Voice Coil Actuator with a flexure design that ensures high alignment ... with cost-effective pricing and is ideally suited where extreme precision is required, such ...
(Date:11/30/2016)... 30, 2016 Biotest Pharmaceuticals Corporation (BPC), a ... announce the addition of its newest plasma collection center ... . The 15,200 square foot state-of-the-art facility officially ... and brings the total number of BPC,s plasma collection ... , BPC,s Chief Executive Officer said "We are pleased ...
(Date:11/30/2016)... DIEGO and BEIJING , ... a leading commercial provider of genomic services and solutions ... announced today that it has completed a USD $75 ... Merchants Bank Co., Ltd.,s CMB International Capital Management ( ... Innovation Investment Management Co., Ltd. ("SDIC Innovation") and Shanghai ...
Breaking Biology Technology:
(Date:11/22/2016)... November 22, 2016 According to the new market ... Print, Face, Vein, Signature, Voice), Multi-Factor), Component (Hardware and Software), Function (Contact ... MarketsandMarkets, the market is expected to grow from USD 10.74 Billion in ... 16.79% between 2016 and 2022. Continue Reading ... ...
(Date:11/21/2016)... , Nov. 21, 2016   Neurotechnology ... object recognition technologies, today announced that the MegaMatcher ... cards was submitted for the NIST Minutiae ... passed all the mandatory steps of the evaluation ... is a continuing test of fingerprint templates used ...
(Date:11/17/2016)... -- Global Market Watch: Primarily supported by ownership ... and Academics) market is to witness a value of US$37.1 ... highest Compounded Annual Growth Rate (CAGR) of 10.75% is foreseen ... period 2014-2020. North America is not ... Europe at 9.56% respectively. Report Focus: ...
Breaking Biology News(10 mins):