SmartRT™ Reverse Transcriptase Kit
SmartRT™ Reverse Transcriptase Kit

SmartRTTM Reverse Transcriptase is an engineered MMLV RT that improves the enzyme’s thermostability, reduces RNase H activity and its cDNA synthesis ability. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized cDNA.  With a 3’ modified oligo dT primer and a 5’  SMART universal oligo containing a terminal complementation to nucleic acids at the 3’  end of the first-strand cDNA, the SmartRTTM reverse transcriptase will produce RACE ready full-length cDNA. 

 
Name Cat# Size Concentration Price Qty
SmartRT™ Reverse Transcriptase
SMRT-100
4000u(40 rxn)
100u/μl
$49.00
SmartRT™ Reverse Transcriptase
SMRT-200
10000u(100 rxn)
100u/μl
$89.00
SmartRT™ Reverse Transcriptase
SMRT-300
40000u(400 rxn)
100u/μl
$299.00
SmartRT™ Reverse Transcriptase
SMRT-OEM
any size
please inquire

General Description

SmartRTTM Reverse Transcriptase is an engineered MMLV RT that improves the enzyme’s thermostability, reduces RNase H activity and its cDNA synthesis ability. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized cDNA.  With a 3’ modified oligo dT primer and a 5’  SMART universal oligo containing a terminal complementation to nucleic acids at the 3’  end of the first-strand cDNA, the SmartRTTM reverse transcriptase will produce RACE ready full-length cDNA. 

Features
MCLAB's reverse transcriptase is one of transcriptases that has the highest thermostability with terminal transferase activity.

Source
E. coli

Applications
Reverse transcription and RACE ready full length cDNA synthesis

Supplied with
5 x first strand cDNA synthesis buffer

Recommended Storage Condition
-20 °C

Unit Definition
One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in 10 minutes at 37°C using poly(A):oligo(dT) as template:primer.

Shipping Condition
dry ice

User protocol
For RACE ready first-strand cDNA Synthesis
Combine the following in separate 0.2-ml PCR tubes (20 µl reaction):
1–9 µl RNA sample
1 µl 5’ SMART universal Primer
1 µl oligo dT Primer
1 µl dNTP
Add sterile H2O to a final volume of 12 µl for each reaction.
Incubate the tubes at 65°C for 5 min
Cool the tubes on ice for 3 min
Add the following to each reaction tube:
4 µl 5X First-Strand Buffer
1 µl DTT (20 mM)
2 µl dNTP Mix (10 mM)
2 µl SmartRT reverse transcriptase
Incubate the tubes:
50°C for 1.5 hr
85°C for 5 min

For regular first-strand cDNA Synthesis

Combine the following in separate 0.2-ml PCR tubes (20 µl reaction):
1–10 µl RNA sample
1 µl oligo dT Primer or Random primer
1 µl dNTP
Add sterile H2O to a final volume of 12 µl for each reaction.
Incubate the tubes at 65°C for 5 min
Cool the tubes on ice for 3 min
Add the following to each reaction tube:
4 µl 5X First-Strand Buffer
1 µl DTT (20 mM)
2 µl dNTP Mix (10 mM)
2 µl SmartRT reverse transcriptase
Incubate the tubes:
50°C for 45 min
85°C for 5 min

SMARTRT-REVERSE
Figure 1.Smart cDNA synthesis compared to conventional cDNA synthesis. Unlike conventional cDNA synthesis methods which involve a multiple enzyme/multiple step procedure, the Smart cDNA synthesis protocol is performed by on reverse transcription reaction, in a single tube, with no adaptor ligation or intervening purification steps. Following PCR amplification, Smart cDNA is immediately available for a variety of downstream applications.

 

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