Navigation Links
Efficient and Reliable Linear,,, Amplification of cRNA

lizing T7 RNA polymerase for the preparation of the labeled target, first described by van Gelder and Eberwine [1], has become a standard procedure (Figure 1 a). This protocol consists of a first- and second-strand cDNA synthesis followed by cRNA in vitro transcription using T7 RNA polymerase. For first-strand cDNA synthesis an oligo(dT) primer linked to the T7 promoter sequence is used. In the next step, this T7 linker allows the linear amplification of the template and the simultaneous incorporation of labeled nucleotides with T7 RNA polymerase, generating singlestranded, labeled cRNA. The complete target-preparation workflow consists of multiple enzymatic steps determining the quality and integrity of the targets.

Experimental Validation

To validate the T7 RNA polymerase-based target synthesis procedure, the Roche Applied Science (RAS) Microarray RNA Target Synthesis Kit (T7) and the kit of a major Supplier (A) were tested on three different human acute myeloid leukemia (AML) research samples and one chronic lymphocytic leukemia (CLL) research sample (Figure 1b).

First, 5 g of total RNA was converted into double-stranded (ds) cDNA using the RAS Microarray cDNA Synthesis Kit (eight independent reactions). One group of ds cDNA samples was purified with the RAS Microarray Target Purification Kit, then amplified with T7 RNA polymerase and labeled with Biotin-UTP, using the RAS Microarray RNA Target Synthesis Kit (T7). The resulting labeled cRNA was purified with the RAS Microarray Target Purification Kit. The ds cDNA of the other sample group was purified according to the Supplier As protocol, followed by in vitro transcription and labeling with Biotin- UTP and Biotin-CTP using Supplier As kit. Purification of the labeled cRNA was carried out accor
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
4. Efficient Transfection of Neurospora Crassa
5. Efficient Transfection of Neurospora Crassa
6. Efficient Recovery of Ultrapure Plasmid DNA
7. Efficient and Reliable PCR Setup Using Eppendorf MasterMix
8. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
9. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
10. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
11. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Efficient and Reliable Linear Amplification cRNA

(Date:9/30/2014)... Md. , Sept. 30, 2014  RegeneRx ... today announced that it has received a Canadian ... by Actin-Sequestering Peptides.  This patent includes claims using ... numerous other actin-sequestering peptides for these purposes.  The ... About RegeneRx Biopharmaceuticals, Inc. ( www.regenerx.com ) ...
(Date:9/30/2014)... Switzerland (PRWEB) September 30, 2014 ... software solutions for drug discovery and related ... FUNGITECT consortium has chosen Genedata Selector as ... sequencing (NGS) data. FUNGITECT is a European ... invasive fungal diseases via individualized anti-fungal drug ...
(Date:9/30/2014)... and ROCKVILLE, Md., Sept. 30, 2014 ... focusing on commercializing market-relevant biohealth innovations and increasing access ... , announced today that venture capitalist, Tania Fernandez ... strategic advisor. Dr. Fernandez will be a member of ... which will provide up to $50 million in seed ...
(Date:9/30/2014)... -- The overall research objective was to capture ... use. Most researchers outsource NGS technologies; however, in-house ... the next 12 months. Data accuracy and operating ... instruments. Disease-related research is the dominant use for ... surge, with half of the respondents planning to ...
Breaking Biology Technology:RegeneRx Receives Canadian Patent for Inhibition or Reversal of Skin Aging 2FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 2FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 3Seasoned Life Sciences Venture Capitalist Tania Fernandez Joins BHI to Help Manage New BioHealth Gap Fund 2Global Next-generation Sequencing Trends 2
... the appointment of James Stoppert as President and Chief Executive Officer ... ... EON) September 3, 2008 -- Segetis is the first and ... wide range of industrial chemicals and plastics that are ultimately derived ...
... China, Sept. 3 ,Acesys Pharmatech and Provid Pharmaceuticals ... expertise and project management,capabilities of Provid in the ... structure of Acesys in China., The combined ... of medicinal chemistry for clients in the pharmaceutical ...
... tested new molecular tools for stem cell research to direct ... development programmes. , A collaborative team of scientists from ... (NESCI) have developed two synthetic molecules which can be used ... into other forms of tissue. , Their use ...
Cached Biology Technology: Stoppert Named as President and CEO at Breakthrough Renewable Technology Firm Segetis, Inc 2Acesys and Provid Announce Alliance for Medicinal Chemistry Services 2New stem cell tools to aid drug development 2New stem cell tools to aid drug development 3
(Date:9/30/2014)... German . ... risky situations such as exposure to predators. Researchers from ... a long-term study on different populations of great tits ... ambient temperature. High metabolic rates and low temperatures were ... birds were more likely to approach potential predators. , ...
(Date:9/30/2014)... Coral reefs face a suite of perilous threats ... coastal development and climate change, fragile coral ecosystems ... Despite this trend, some species of corals surrounding ... a natural protector in their tropical environment: coral ... Natural History,s Smithsonian Marine Station scientist Seabird McKeon ...
(Date:9/30/2014)... -- Winners of The Economist,s prestigious Innovation awards 2014 ... place at the JW Marriott Hong Kong on Friday, October 10 ... their experiences and the lessons those hold for others. The awards, ... the past decade, will be presented at a ceremony in ... will be the first time the ceremony has come to ...
Breaking Biology News(10 mins):Risky metabolism 2Smithsonian scientists discover coral's best defender against an army of sea stars 2Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 2Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 3Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 4Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 5Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 6Hear from The Economist's 2014 Innovation Award Winners at Innovation Summit on October 10th in Hong Kong 7
... like AND and OR Boolean logic gates, producing an output ... in the May issue of Nature Chemical Biology , ... tiny circuits. Study leader Takanari Inoue, Ph.D., an assistant ... the Institute of Basic Biomedical Sciences, Center for Cell Dynamics ...
... of published studies on the impact of bark beetles ... more complete picture of the effect of this destructive ... a wide variety ofand often contradictoryconclusions about the bark ... researchers from the USDA Forest Service,s Pacific Southwest Research ...
... Calif.) Synthetic platelets have been developed by UC ... Research Institute and Sanford-Burnham Institute in La Jolla, Calif. ... Materials in a paper titled "Platelet Mimetic Particles ... the components of blood that allow it to prevent ...
Cached Biology News:Training cells to perform Boolean functions? It's logical 2Training cells to perform Boolean functions? It's logical 3New report examines effects of trees killed by bark beetles on wildfire 2UC Santa Barbara researchers develop synthetic platelets 2
... Motic's B3 series of microscopes offer ... microscope required for laboratory work. Equipped with ... Super Contrast (ASC) objectives. The B3 is ... The Siedentopf binocular tube eliminates the need ...
... SYBR Green II RNA gel ... gel stain that has bright fluorescence when ... gels, making it ideal for use with ... scanners or standard UV transilluminators. A 500 ...
... SYBR Green II RNA gel stain ... has bright fluorescence when bound to RNA and ... use with either formaldehyde/agarose or polyacrylamide gels using ... stains 100 minigels. A 500 µL unit size ...
... Glutathion-S-transferases (GST) are a group ... the detoxification processes. The GSTs are ... Pi, Teta and microsomal. The classes ... and s Sample Size: ...
Biology Products: