商品编号


DS010910



品牌


Lucigen



公司


Lucigen



公司分类


Nuclease Resistant In Vitro Transcription Kits



Size

10 Reactions





商品信息

Produce fully RNase A-resistant RNA that is perfectly suited for RNA aptamer synthesis, as well as antisense RNA and RNA interference (RNAi) experiments.

Protect transcribed RNA from RNAse degradation

Produce high RNA yields

Select and optimize RNA aptamers using SELEX screening

See FAQ




The DuraScribe? T7 Transcription Kit* is an
in vitro
transcription kit that produces RNA - called DuraScript? RNA - that is completely resistant to RNase A digestion.

The DuraScribe T7 Transcription Kit includes DuraScribe T7 RNA Polymerase, an enhanced formulation of the Y639F mutant
1
of T7 RNA Polymerase that efficiently incorporates 2?-Fluorine-CTP (2?-F-dCTP), 2?-Fluorine-UTP (2?-F-dUTP), ATP and GTP into RNase-resistant RNA transcripts called DuraScript RNA (Figure 1). The DuraScribe T7 RNA Polymerase uses the same T7 promoters as the wild-type T7 RNA Polymerase.

Applications

DuraScript RNA has demonstrated advantages for RNA aptamer synthesis, and ribozyme, RNA interference (RNAi) and antisense RNA studies.

Features

DuraScript RNA is resistant to Rnase A and Dnase

High Yields of DuraScript RNA

T7
in vitro
transcription reations.
DuraScribe T7 RNA Polymerase uses the same T7 promoters as the wild type T7 RNA Polymerase.

Ideal for RNA aptamer studies
. Numerous citations using the DuraScribe Kit with RNA aptamers

SELEX-compat
IBL
e
. DuraScript RNA can be used in SELEX selection processes.

Make long or short DuraScript RNA
. Double-strand oligos, linearized plasmids, and PCR products with a T7 polymerase promoter can be transcribed.

Table 1. Yield of DuraScript? RNA from a DuraScribe? Kit reaction
. One microgram of a 3-Kb DNA template was linearized at different sites and then transcribed in a DuraScribe T7 Transcription Kit reaction for 4 hours. The yield of DuraScript RNA produced from each template is shown in micrograms (?g) and in picomoles (pmol).

Size of DuraScript RNA produced

DuraScript RNA Yield (?g)

DuraScript RNA Yield (pmol)

2600 nts

100 ?g

116 pmol

1400 nts

58 ?g

124 pmol

330 nts

18 ?g

164 pmol

88 nts

9 ?g

307 pmol


?





Figure 1 (click to enlarge). The DuraScribe? T7 RNA Polymerase efficiently incorporates 2?-F-dCTP and 2?-F-dUTP into full-length DuraScript? RNA
. The presence of the fluorine at the 2?-position of the 2?-F-dC and 2?-F-dU nucleotides prevents digestion by Rnase A.




Figure 2. Yield of RNA from a DuraScribe? T7 Transcription reaction.
A standard reaction (4-6 hours) produced 40-60 ?g of a 1.4-kb DuraScript? RNA.




Figure 3. DuraScript? RNA is resistant to Rnase A digestion.
A 1.4-kb standard RNA transcript and a 1.4-kb DuraScript RNA transcript were each incubated with 1 U of highly purified Rnase A for 30 minutes. The standard RNA transcript was completely degraded while the DuraScript RNA transcript remained intact. Lane M, size ladder; lane 1, 1.4-kb standard RNA transcript; lane 2, standard RNA after Rnase A treatment; lane 3, 1.4-kb DuraScript RNA; lane 4, DuraScript RNA after Rnase A treatment.




Figure 4. DuraScript? RNA is completely resistant to "finger" nucleases.
A 1.4-kb standard RNA transcript and a 1.4-kb DuraScript RNA transcript were produced using sterile water or water that had been contaminated by exposure to the hands of a test subject. The standard RNA shows extensive degradation from "finger" nucleases in the contaminated water while the DuraScript RNA remains fully intact. M, Size ladder; Lane 1, Standard RNA transcript; Lane 2, Standard RNA after "finger" nuclease exposure; Lane 3, DuraScript RNA; Lane 4, DuraScript RNA after "finger" nuclease exposure.


* Covered by issued and/or pending patents.

References

Sousa, R. and P
ADI
lla, R. (1995) EMBOJ. 14:18 4609-4621.


ORDER INFORMATION
Contents: DuraScribe T7 Enzyme Mix, RNase-Free DuraScribe T7 10X Reaction Buffer, ATP, GTP, 2-F-dCTP, 2-F-dUTP, DNase I, DTT, Control Template DNA (linearized), Sterile Deionized Water.