商品编号


SYG421010



品牌


Lucigen



公司


Lucigen



公司分类


Reverse Transcriptase Enzymes



Size

10 rxn





商品信息

A RNase H- reverse trancriptase with improved thermal st
ABI
lity, efficiency and processivity for the production of
CDN
A from long and difficult mRNA templates.


Improved thermal st
ABI
lity
: Active up to 85?C

High yields of first strand
CDN
A

High processivity:
Produces
CDN
A of longest mRNAs in the mammalian transcriptome

Great sensitivity:
? Works with pg amounts of starting total RNA

High reproducibility

Robust and reliable





SunScript? Reverse Transcriptase RNaseH- is a engineered reverse transcriptase based on the well characterized HIV-1 RT with high
Thermo
st
ABI
lity and no Rnase H activity. It has RNA- and DNA-dependent polymerase activity. This enzyme is active up to 85°C. The elimination of RNase H activity enables this enzyme to produce high yields of full length
CDN
A from long RNA molecules, and the high thermal st
ABI
lity promotes reverse transription through complex secondary structures.



Reverse transciption at temperatures as high as 85°C


High sensitivity over a wide RNA of starting RNA amounts

Reverse transcription of long mRNAs

High
CDN
A synthesis rate



Reverse transciption at temperatures as high as 85°C


100 ng of total RNA from HeLa cells were reverse transcribed at the indicated temperatures for 60 min with SunScript Reverse Transcriptases and Oligo(dT)18?primer, followed by inactivation at 95°C for 10 min. Presence of GAPDH
CDN
A was detected by PCR (product length 305 bp).

High reaction temperatures are advantageous to transcribe RNAs with complicated secondary structures.

?

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High sensitivity over a wide range of RNA concentrations


Sensitivity test: RT reactions were performed at 70°C for 60 min. Inactivation at 95°C for 10 min.

Primer set : Human GAPDH305bp set

1- 1 ?g Human total RNA from HeLa cells
2- 100 ng Human total RNA from HeLa cells
3- 10 ng Human total RNA from HeLa cells
4- 1 ng Human total RNA from HeLa cells
5- 100 pg Human total RNA from HeLa cells

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Transcription of long mRNAs


1- SunScript? RT RNaseH+ replicate 1
2- SunScript? RT RNaseH+ replicate 2
3- SunScript? RT RNaseH+ replicate 3
4- SunScript? RT RNaseH- replicate 1
5- SunScript? RT RNaseH- replicate 2
6- SunScript? RT RNaseH- replicate 3

Assay for long mRNA reverse transcription: 3 independent RT reactions were performed at 65°C for 60 min using Oligo(dT)20?primers. Inactivation at 95°C for 10 min. PCR Primer sets : mm
NEB
(>16 kb from 3’end of mRNA; mRNA total length 22,5 kb); mmDnah8 (> 8,5kb from 3’end of mRNA; mRNA total length 14,5 kb)

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High
CDN
A synthesis rate


1 ?g of total RNA from HeLa cells was reverse transcribed at 65°C for the indicated times with SunScript? Reverse Transcriptases and Oligo(dT)18 primer, followed by inactivation at 95°C 10 min.

PCR Primer set: hsHERC4033 (>11 kb from 3’end of mRNA; mRNA total length 15.3 kb)


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ORDER INFORMATION
Each SunScript? RT RNase H- contains: enzyme, 5X Reaction Buffer and DTT. The complete manual is available under the Manual tab.
Lucigen
is an authorized distributor of Sygnis products in the US.

SunScript? Reverse Transcriptase RNaseH- is intended for molecular
BIOLOG
y use only and
in vitro
use only. This product is not intended for diagnosis, prevention or treatment of a disease in human beings or animals.