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Reverse Transcriptases

Home > PRODUCTS > Reverse Transcriptases

SuperiorScript III cDNA Synthesis Kit new

No Product Name Cat.# Conc Size Price(₩) Add
1 SuperiorScript III cDNA Synthesis Kit EZ405S 200 unit/μl 50 rxns ₩ 390,000 추가
2 SuperiorScript III cDNA Synthesis Kit EZ405M 200 unit/μl 100 rxns ₩ 702,000 추가
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Product description

SuperiorScript III Reverse Transcriptase is genetically engineered version of M-MLV RT with reduced RNase H activity and increased thermostability, thus this enzyme can synthesize first-strand cDNA at elevated temperatures up to 55 . SuperiorScript III Reverse Transcriptase shows improved thermostability and reduced RNase H activity than the competitor’s enzyme. This enzyme is capable of synthesizing first strand cDNA at 42~55 with high specificity and yield. Synthesis of longer products (12 kb or more) are possible than the previous versions.

 

Characteristics

- Produces excellent yields of amplifiable cDNA

- Can synthesize long cDNA targets, up to 12 kb, from small amounts of input RNA

- Able to reverse transcribe through RNA secondary structures

- Strict enzyme purity specifications

 

Applications

- cDNA cloning or cDNA library preparation

- Two step RT-PCR (end-point or real-time quantitative PCR)

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

- Inhibitor-free

- Satisfactory yield and length of cDNA products

 

RNase H digestion

- RNase H can improve PCR sensitivity especially on the long template after synthesizing first-strand cDNA by

  eliminating RNA template on cDNA:RNA hybrid.  RNase H which is component of this kit can be properly 

  used if necessary.

- The existence of RNase H while synthesizing the first-strand cDNA could decrease the yield of full-length 

  cDNA.

 

Standard reaction conditions

'Standard conditions' is only a general recommendation. The experimental conditions should be adjusted according to the purpose and sample.

 

Standard reaction conditions

10X RT Buffer

2 μl

SuperiorScript III Reverse Transcriptase (200 units/μl)

1 μl

dNTP Mixture (10 mM each)

1 μl

a)Template RNA

X μl

b)Primer

1 μl

0.1 M DTT

2 μl

RNase Inhibitor (40 units/μl)

0.5 μl

Sterile water (RNase free)

up to 20 μl

a)Prepare one of the following RNA template.

    - Total RNA: 1 pg~5 μg

    - Messenger RNA (mRNA): 1 pg~500 ng

b)Prepare one of the following primers.

    - Oligo (dT)18: 50 μM~100 μM

    - Random hexamer: 50 μM~100 μM

    - Specific primer: 15 pmol~20 pmol

→ An additional annealing step is recommended:

    ∙ If using random hexamer, incubate at 25 for 10 min.

→ Incubate at 50 for 50 min.

→ Incubate at 85 for 5 min to inactivate the reaction.

If using this kit for the first time, highly recommend visiting our website (www.enzynomics.com) to read the detailed version of protocol.

 

Note

SperiorScript III Reverse Transcriptase performs optionally over the full range of 42~55. Typically, 50 is a good starting point. For RNAs containing secondary structure and other challenging targets, a synthesis temperature of 55 may be used without loss of performance.

 



Figure 1. SuperiorScript III cDNA Synthesis Kit shows comparable efficiency and sensitivity on real-time PCR with product from company I.

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