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SuperiorScript II Reverse Transcriptase

No Product Name Cat.# Conc Size Price(₩) Add
1 SuperiorScript II Reverse Transcriptase RT005M 200 units/μl 10,000 units ₩ 144,000 추가
2 SuperiorScript II Reverse Transcriptase RT005L 200 units/μl 50,000 units ₩ 576,000 추가
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Product description

SuperiorScript II Reverse Transcriptase is genetically engineered version of M-MLV RT which is reduced RNase H activity and increased thermostability, thus this enzyme can synthesize cDNA at high temperatures more than M-MLV RT. SuperiorScript II Reverse Transcriptase shows improved cDNA yields and RNA detection sensitivity. This enzyme is capable of synthesizing cDNA longer than 12.3 kb from messenger RNA.

 

Applications

- Synthesis of first-strand cDNA,

- Array labeling

- cDNA library construction

- 3’ and 5’ RACE, RT-PCR

- Primer extension

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

- Inhibitor-free

- Satisfactory yield and length of cDNA products

 

Unit definition

One unit is the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble materials using 0.4 mM poly(rA)-oligo(dT) as substrate at 37 in 10 min.

 

Supplied with

- SuperiorScript II Reverse Transcriptase

- 5X First-Strand buffer

- dNTP Mixture (10 mM each)

- 0.1 M DTT

- Sterile water (RNase free)

 

Storage buffer

20 mM Tris-Cl (pH7.5), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.01% NP40, 50% glycerol

 

5X First-Strand buffer

250mM Tris-HCl (pH 8.3), 375 mM KCl, 15mM MgCl2

 

Standard reaction conditions

5X First-Strand buffer

4 μl

SuperiorScript II Reverse Transcriptase (200 units/μl)

1 μl

dNTP Mixture (10 mM each)

1 μl

0.1 M DTT

2 μl

a)Template RNA

X μl

b)Primer

1 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 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

→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.

→Incubate at 42 for 60 min.

→Incubate at 70 for 15 min to inactivate the reaction.




Figure 1. Thermostability of SuperiorScript II on 3 targets

cDNA was synthesized from 500 ng total HeLa RNA for beta-actin 184bp, GAPDH 395 bp and TFR 738 bp. One-tenth of the cDNA reaction was used for 30 cycles of PCR with nTaq-multi HOT (Cat.# P725HC).





Figure 2. Sensitivity of SuperiorScript II

cDNA was Synthesized from HeLa total RNA with oligo(dT) primer using buffer supplied with each RT and recommended conditions. 10% of cDNA reaction was added to 20 μPCR reaction containing primers for 500bp GAPDH gene and 2X TOPsimple™ DyeMIX-HOT (Cat.# P510H), 30 cycle1 min/kb.




Figure 3. High sensitivity using SuperiorScript II for RT-qPCR

cDNA was synthesized using SuperiorScript II from 100 ng to 0.1 ng total HeLa RNA for beta-actin 117 bpcDNA was amplified using TOPreal™ qPCR 2X PreMIX (SYBR Green with low ROX) (Cat.# RT500) on the Bio-Rad CFX-96.

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