商品编号


SMRT-OEM



品牌


MCLab



公司


MCLAB.Inc



公司分类


Reverse Transcriptase and RNA Polymerases



Size

any size

商品信息

SmartRT
TM
Reverse Transcriptase is an engineered MMLV RT that improves the enzyme’s
Thermo
st
ABI
lity, reduces RNase H activity and its
CDN
A synthesis
ABI
lity. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized
CDN
A. ?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
CDN
A, the SmartRT
TM
reverse transcriptase will produce RACE ready full-length
CDN
A.?




General Description

SmartRT
TM
Reverse Transcriptase is an engineered MMLV RT that improves the enzyme’s
Thermo
st
ABI
lity, reduces RNase H activity and its
CDN
A synthesis
ABI
lity. The enzyme also has a terminal transferase activity, where it adds a few extra nucleotides to the end of the synthesized
CDN
A. ?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
CDN
A, the SmartRT
TM
reverse transcriptase will produce RACE ready full-length
CDN
A.?


Features

MCLAB
's reverse transcriptase is one of transcriptases that has the highest
Thermo
st
ABI
lity with terminal transferase activity.


Source
E. coli


Applications
Reverse transcription and RACE ready full length
CDN
A synthesis


Supplied with
5 x first strand
CDN
A 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
CDN
A Synthesis
Combine the following in separate 0.2-ml PCR tubes (20 ?l reaction):
1–9 ?l RNA sample
1 ?l 5’ SMART universal Primer
1 ?l oligo dT Primer
1 ?l dNTP
Add sterile H
2
O to a final volume of 12 ?l 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 ?l 5X First-Strand Buffer
1 ?l DTT (20 mM)
2 ?l dNTP Mix (10 mM)
2 ?l SmartRT reverse transcriptase
Incubate the tubes:
50°C for 1.5 hr
85°C for 5 min
For regular first-strand
CDN
A Synthesis
Combine the following in separate 0.2-ml PCR tubes (20 ?l reaction):
1–10 ?l RNA sample
1 ?l oligo dT Primer or Random primer
1 ?l dNTP
Add sterile H
2
O to a final volume of 12 ?l 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 ?l 5X First-Strand Buffer
1 ?l DTT (20 mM)
2 ?l dNTP Mix (10 mM)
2 ?l SmartRT reverse transcriptase
Incubate the tubes:
50°C for 45 min
85°C for 5 min