Navigation Links
Amino Allyl Labeling for Array Analysis


RNA is often labeled with fluorescent dyes (i.e. Cy3 or Cy5) for microarray analysis. Label may be directly incorporated during cDNA synthesis, aRNA amplification or a post-aRNA reverse transcription (RT) reaction, or attached via a chemical coupling reaction performed after any enzymatic reactions (post-synthesis). Direct labeling requires the enzymatic incorporation of a Cy-labeled dNTP/NTP into the cDNA, aRNA or RT product. Alternatively, indirect labeling usually incorporates an intermediary nucleotide to which the fluorescent label is later attached. Although seemingly more convenient, direct labeling can be problematic for a number of reasons. Cy-labeled dNTPs/NTPs are not incorporated by reverse transcriptase or RNA polymerases with the same efficiency as unlabeled NTPs, leading to variable incorporation efficiency of different dyes (e.g. Cy3-NTP vs. Cy5-NTP). In addition, to be effective, direct incorporation often requires the use of more expensive modified nucleotides.

Post-synthesis labeling avoids the problems inherent to direct enzymatic incorporation of Cy-labeled dNTPs/NTPs. With indirect labeling, amine-modified NTPs (amino allyl-dUTP/UTP) are incorporated during reverse transcription or aRNA amplification. Amino allyl-NTPs are incorporated with similar efficiency as unmodified NTPs during enzymatic reactions such as reverse transcription. Coupling of Cy3 or Cy5 dyes to the amino allyl cDNA or aRNA, post-synthesis, results in probe populations with equivalent labeling efficiencies.

Amino allyl-dUTP contains a reactive amino group on a 2-carbon spacer attached to the methyl group on the base portion of dUTP. During the coupling reaction, after cDNA or aRNA synthesis, this amino group reacts with the NHS ester of a monoreactive Cy dye.

Ambion has several products for the post-synthesis labeling of nucleic acids for array analysis. The Amino Allyl cDNA Labeling Kit contains all the necessary reagents (except monoreactive Cy dyes) for the synthesis of amino allyl labeled cDNA by reverse transcription and subsequent Cy dye coupling. The MessageAmp aRNA Kit generates amplified aRNA for array analysis. Amino allyl UTP can be incorporated into the transcription step of this procedure to produce amino allyl labeled RNA ready for Cy dye coupling. Finally, amino allyl dUTP/UTP, available separately from Ambion, can be incorporated into any reverse transcription reaction.


Cy3 and Cy5 monoreactive dyes are available for Amersham Biosciences.
Cy3 and Cy5 are trademarks of Amersham Biosciences.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
4. New Microarray Labeling Kit for Preparing cDNA Probes
5. aRNA Synthesis and Labeling for Array Analysis
6. Fluorescently Labeling Long dsRNA with the Silencer siRNA Labeling Kit
7. Efficient Europium Labeling of Lymphokine-Activated Killer Cells Using the Gene Pulser II with RF Module, Rev C
8. USB OptiKinase Eliminates Base Bias in Labeling of Oligonucleotide 5-Ends
9. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
10. Human Universal cDNA Library Array I
11. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Amino Allyl Labeling for Array Analysis

(Date:10/1/2014)... , October 1, 2014 ... to software invited to engage around the theme, ,Commercial ...     Elsevier , ... products and services, will host a gathering of some ... issues and trends impacting healthcare today and in the ...
(Date:9/30/2014)... a well-known optical phenomenon called thin-film interference, a ... the ability to "paint" ultra-thin coatings onto a ... making future, flexible electronic devices, creating advanced solar ... ships and spacecraft with extremely lightweight decorative logos. ... it becomes iridescent, revealing a myriad of colors ...
(Date:9/30/2014)... analog of one of nature,s most efficient light-harvesting ... led by Alejandro Briseno of the University of ... developing long-sought polymer architecture to boost power-conversion efficiency ... devices. , Briseno, with colleagues and graduate ... University and Dresden University of Technology, Germany, report ...
(Date:9/30/2014)... Sept. 30, 2014 Report Details ... highest potential revenues Do you want to find ... you revenue predictions for those biological drugs from ... you find financial data, R&D trends, opportunities and ... sales forecasts to 2024 at overall world market, ...
Breaking Biology Technology:Elsevier Healthcare and Pharmaceutical Economics Conference 2014 To Convene Industry Leaders to Discuss Top Healthcare Business Issues 2Elsevier Healthcare and Pharmaceutical Economics Conference 2014 To Convene Industry Leaders to Discuss Top Healthcare Business Issues 3Taking thin films to the extreme 2Blades of grass inspire advance in organic solar cells 2Blades of grass inspire advance in organic solar cells 3Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 2Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 3Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 4Biosimilars and Follow-On Biologics: World Industry and Market Prospects 2014-2024 5
... Madison, Wis. The nation's dysfunctional healthcare system may ... adoption of electronic health records, according to a featured speaker ... Jeff Grossman Dr. Jeff Grossman, president and CEO ... perspective on the role of IT in healthcare delivery. As ...
... web site in your industry is a noble goal, a far ... ,The best means of measuring your web site's popularity is by ... of other web sites linking to it. Each time another web ... of confidence in your site and the quality of its content. ...
... England - A British court has ruled in favor of ... Apple Corps, Ltd. , a company the Beatles established in the ... that Apple Computer, Inc. is entitled to use the ... by Apple Corps. The dispute, which centered on a 1991 agreement, ...
Cached Biology Technology:U.S. health system could undermine EHR progress 2U.S. health system could undermine EHR progress 3Link Popularity: The Internet's 'In' Crowd 2Link Popularity: The Internet's 'In' Crowd 3Apple Computer may use Apple logo on iTunes 2
(Date:9/30/2014)... National Institutes of Health (NIH) awarded Lawrence Livermore ... an electrode array system that will enable researchers ... unprecedented resolution and scale. , LLNL,s grant-funded project ... Obama,s BRAIN (Brain Research through Advancing Innovative ... revolutionize our understanding of the human mind and ...
(Date:9/30/2014)... drinking water may increase the risk of stillbirth ... led by a Boston University School of Public ... journal Environmental Health , compared 1,091 PCE-exposed ... in Cape Cod, Ma., where water was contaminated ... by the installation of vinyl-lined asbestos cement pipes. ...
(Date:9/30/2014)... FRANKFURT. Truffles, along ... world. Because they grow underground, people use trained dogs or ... is not only of interest to gourmets. A group of ... University Frankfurt have discovered that the smell of white truffles ... truffle fruiting bodies. , White truffles from the Piedmont region ...
Breaking Biology News(10 mins):NIH taps lab to develop sophisticated electrode array system to monitor brain act 2NIH taps lab to develop sophisticated electrode array system to monitor brain act 3Contaminated water linked to pregnancy complications, BU study finds 2On the trail of the truffle flavor 2
... 30, 2013) La Jolla Institute for Allergy ... a wholly-owned subsidiary of Kyowa Hakko Kirin Co., Ltd. ... the signing of a new agreement that marks another ... in the world. The six-year agreement continues a ...
... presented at the Annual Meeting of the Society for the ... into all aspects of eating and drinking behavior, shows that ... at risk to develop obesity and type 2 diabetes. ... stress exposure and passive stress coping style might predispose offspring ...
... be presented at the Annual Meeting of the Society for ... research into all aspects of eating and drinking behavior, found ... changes appear to be independent of taste nerve activity. ... and it is generally believed that reduced consumption is related ...
Cached Biology News:La Jolla Institute continues longtime collaboration with Kyowa Hakko Kirin California 2La Jolla Institute continues longtime collaboration with Kyowa Hakko Kirin California 3Taste preference changes in different life stages of rats 2
Melanoma Antigen, Family C, 1 (MAGEC1) Rabbit anti-Human Polyclonal Antibody Immunogen: Synthetic peptide - KLH conjugated Family: New Recommended Storage: Long term: -20C; Short term: +4...
X box-binding protein 1...
Mouse monoclonal [AE3] to Cytokeratin 1 - 8, prediluted ( Abpromise for all tested applications). Antigen: Tissue / cell preparation - Human epidermal cells. Entrez GeneID: 140807 Swiss P...
Mouse HAI-1 Ectodomain MAb (Clone 199732)...
Biology Products: