Navigation Links
ArrayScript RT: Maximize Yields of Full-length cDNA


Get Full-length cDNA
Ambion scientists have engineered a mutant form of M-MLV reverse transcriptase to improve production of full-length cDNA. This engineered reverse transcriptase, ArrayScript (patent pending), produces significantly higher yields of full-length cDNA than wild type RTs, and rivals even the newest engineered RTs on the market (Figure 1). Efficient synthesis of full-length cDNA is important for microarray analysis; it results in low 3'/5' ratios, which are a hallmark of high quality microarray data.

Figure 1. ArrayScript Reverse Transcriptase Synthesizes Intact cDNA Products as Long as 9 kb. cDNA synthesis was performed with 100 U of either ArrayScript RT, SuperScript II (SSII), SuperScript III (SSIII), or wild type (wt) M-MLV RT in a 20 l reaction volume containing a synthetic, poly(A)-tailed 9 kb transcript (1 g), 1X RT buffer, 0.5 M oligo(dT) primer, 0.5 mM dNTPs, 40 U RNase Inhibitor, and 0.1 l [a-32P]dATP. Reactions were incubated for 1 hr at 42C and quenched with EDTA. The cDNA products were resolved on a 1% denaturing agarose gel and detected with a phosphorimager.


Double the Yield in RNA Amplification
One of the most rigorous applications for an RT enzyme is RNA amplification from low amounts of total RNA. In this application, first-strand cDNA generated with ArrayScript resulted in up to twice the yield of amplified RNA compared to cDNA synthesized using wild type M-MLV, AMV, or other engineered M-MLV reverse transcriptases (Figure 2). When starting with low amounts of total RNA, ArrayScript often eliminates the need for two rounds of amplification, still generating sufficient cRNA for array analysis. This translates to significant savings of time and reagents.

Figure 2. ArrayScript Reverse Transcriptase Produces up to Twice the cRNA Yield in MessageAmp II. A comparison of ArrayScript RT (AS; 200 U), SuperScript II (SSII; 200 U); wild type M-MLV RT (200 U), and wild type AMV RT (10 U) in MessageAmp II conditions demonstrates that ArrayScript outperforms other commercial RTs. 90 ng HeLa-S3 total RNA (28S/18S ratio = 1.7) was heat denatured in the presence of T7 Oligo(dT) at 70C for 10 minutes. Reverse transcription was initiated with the RT enzyme and incubated at 42C for 2 hr in an air incubator. Following first-strand cDNA synthesis, DNA polymerase was added for second-strand synthesis, which was incubated at 16C for 2 hr in a thermocycler (heated lid option turned off). The double-stranded cDNA was then filter purified and added to a 4 hr in vitro transcription reaction. Last, the resulting cRNA was purified by glass filter and eluted in nuclease-free water for analysis by Agilent 2100 bioanalyzer and NanoDrop Spectrophotometer.


ArrayScript is Suitable for all RT Applications
Reverse transcriptases are also used in RT-PCR, cDNA library construction, and RLM-RACE. In most of these applications, synthesis of full-length cDNA is a critical parameter that affects final experimental results. For example, high yields of full-length cDNA result in the generation of cDNA libraries that more faithfully represent the cellular transcriptome. The higher yields possible with ArrayScript also make it an excellent choice for general reverse transcriptase applications that are not length sensitive, such as real-time RT-PCR.

ArrayScript is provided in two convenient sizes, 4,000 U and 10,000 U, and a 10X Reaction Buffer is provided with the enzyme.

ArrayScript is a critical component of Ambions MessageAmp II aRNA Amplification Kit. Learn how this kit outperforms other amplification products.


back to top


Ordering Information Cat# Product Name Size 2048 ArrayScript 4000 U 2049 ArrayScript 10,000 U
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Maximize Viral RNA Yield from Biological Fluid
2. Rapid Chromatography Optimization by pH Scouting Using the New BioLogic Maximizer Valve System, Rev A
3. Yeast Protein Production System Features High Yields and One-Step Purification
4. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
5. Synthesize High Yields of Biotinylated aRNA
6. Translate High Yields of Active Protein
7. Translate High Yields of Biologically Active Proteins
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:3/29/2017)... ... 29, 2017 , ... ACEA Biosciences, a privately owned biotechnology ... award to noteworthy scientists who will be presenting research using ACEA Biosciences’ technology ... awards are being given to two postdoctoral fellows studying pathogen capture by neutrophils, ...
(Date:3/29/2017)... 29, 2017 Research and Markets has announced ... Trends - Industry Forecast to 2025" report to their offering. ... The Global ... around 7.8% over the next decade to reach approximately $825.4 million ... estimates and forecasts for all the given segments on global as ...
(Date:3/29/2017)... TORONTO , March 29, 2017 /PRNewswire/ -  GeneNews Limited ... of BreastSentry™ , a new risk stratification test for ... reference lab, Innovative Diagnostics Laboratory ("IDL"). BreastSentry incorporates a blood-based ... five-year and lifetime risk for developing breast cancer.   ... BreastSentry measures ...
(Date:3/28/2017)... ... March 28, 2017 , ... ... Association for the Advancement of Science (AAAS), the world's largest general scientific ... enhance its high-impact scholarly collection across its cross-platform reference management system. , ...
Breaking Biology Technology:
(Date:3/24/2017)... 2017 Research and Markets has announced the ... & Trends - Industry Forecast to 2025" report to their ... The Global ... CAGR of around 15.1% over the next decade to reach approximately ... the market estimates and forecasts for all the given segments on ...
(Date:3/22/2017)... March 21, 2017 Vigilant Solutions , ... law enforcement agencies, announced today the appointment of retired ... of public safety business development. Mr. Sheridan ... experience, including a focus on the aviation transportation sector, ... recent position, Mr. Sheridan served as the Aviation Liaison ...
(Date:3/9/2017)... , Australia , March 9, ... study data at the prestigious World Lung Imaging Workshop ... Andreas Fouras , was invited to deliver the ... pulmonary medicine. This globally recognised event brings together leaders ... share the latest developments in lung imaging. ...
Breaking Biology News(10 mins):