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TOPscript™ One-step RT PCR Kit

No Product Name Cat.# Conc Size Price(₩) Add
1 TOPscript™ One-step RT PCR Kit RT410S - 100 reactions ₩ 210,000 추가
2 TOPscript™ One-step RT PCR Kit RT410M - 200 reactions ₩ 378,000 추가
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Product description

TOPscript One-step RT PCR Kit is ready-to-use PCR mixture containing 4X concentration of components required for RT-PCR in one tube. For reactions, simply add template RNA and a pair of primers resuspended in water. Reverse Transcriptase is an improved version of M-MLV Reverse Transcriptase. It lacks RNaseH activity, and is highly heat stable. Therefore, this product is most suitable to efficiently synthesizing long cDNA by incubating reactions at elevated (50) temperature since high temperature helps removal of secondary structure present in long RNA transcripts. In addition, HOT start Taq DNA polymerase is a derivative of Taq polymerase which is greatly improved in both fidelity and yield for polymerase chain reaction.This become more prominent as cDNA to be synthesized is long. In addition, it has a wider range of optimal concentrations of primers.

 

Characteristics

- Single-tube convenience

- Synthesis of long cDNA

- Excellent sensitivity

 

Applications

- Virus detection

- Single-cell RT-PCR

- Gene-expression analysis

- cDNA library construction

- 3 and 5 RACE, RT-PCR

 

Components (4X)

- Reverse Transcriptase

- Hot-start Taq DNA Polymerase

- dNTP mixture

- One-step RT PCR reaction buffer

- Stabilizer

 

Storage

Stable up to 6 months at -20 (Storage at -20 is rcommended).

 

Standard reaction conditions

- a)RT-PCR mixture

TOPscript™ One-step RT PCR kit

5 μl

Template (total RNA, 10 ng ~ 1 μg/μl)

X μl

Gene specific primers (2~20 pmol/μl)

1 μl

Sterile water

up to 20 μl

aAssemble the reaction mixture on ice

 

- RT-PCR cycle

Reverse Transcription

50~55

30 min

PCR reaction

Initial denaturation

95

10 min

Denaturation

95

5 sec

b)Annealing

55~65

30~60 sec

Annealing & Elongation

60

30 sec

Number of cycles

30~45 times

When cycles are over, keep the reaction mixture at 4; may add 10 mM EDTA until use to prevent DNA degradation.

b)Recommended annealing temperatures is 5 to 10 below the lower Tm of the two primers used.


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