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Reverse Transcriptases

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TOPscript™ RT DryMIX

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPscript™ RT DryMIX RT100 - 96 tubes ₩ 210,000 추가
2 TOPscript™ RT DryMIX RT101 - 480 tubes ₩ 840,000 추가
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Product description

TOPscript™ RT DryMIX is formulated for maximal stability of TOPscript™ Reverse Transcriptase that is dried with reaction Buffer, dNTP mixture, and stabilizer. This product is ready to use upon the addition of primers and template RNA in distilled water to each tube. TOPscript™ Reverse Transcriptase is genetically engineered version of M-MLV RT which is highly thermostable, thus can synthesize cDNA at elevated temperatures up to 60 This property is very useful when RNA templates are long and have extensive secondary structures. TOPscript™ Reverse Transcriptase is capable of synthesizing cDNA longer than 20 kb from messenger RNA.

 

Characteristics

- Simplicity: The dried pellet in TOPscript™ RT DryMIX is readily soluble upon addition of

primers and template RNA in water.

- Handling: Well-suited to analysis of a large number of samples.

- Stability: The presence of a unique stabilizer ensures great stability of TOPscript™ Reverse Transcriptase.

- Synthesis of long cDNA

- Excellent sensitivity

 

Applications

- Synthesize first-strand cDNA,

- Array labeling

- cDNA libraries

- 3′and 5′RACE, RT-PCR

- Primer extension

 

Components

- TOPscript Reverse Transcriptase

- TOPscript RT Buffer

- dNTP mixture

- RNase inhibitor

- Stabilizer

 

Storage buffer

20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 1 mM DTT, 0.1 mM, EDTA, 0.01% NP40, 50% glycerol

 

Standard reaction conditions

TOPscript™ RT DryMIX

1 tube

a)Template RNA

X μl

b)Primer

1 μl

Sterile water (RNase free)

up to 20 μl

a)Prepare one of the following RNA template.

    - Total RNA: 1 ng~5 μg

    - Messenger RNA (mRNA): 1 ng~250 ng

    - Specific RNA: 0.01 pg~0.5 μg

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

→ Dissolve the dried pellet by vortexing and briefly spin down.

→ An additional annealing step is recommended:

    - if using oligo(dT)18, incubate at 42 for 5 min.

    - if using random hexamer, incubate at 25 for 10 min.

→ lncubate at 42~60 for 60 min.

→ lncubate at 95 for 5 min to inactivate the reaction.

 

Note

TOPscript™ Reverse Transcriptase performs optionally over the full range of 42~60. Typically, 50 is a good starting point. For RNAs containing secondary structure and other challenging targets, a synthesis temperature of 60 may be used without loss of performance.

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