RNA Polymerase T7
更新时间:2016-12-01 10:48:29 信息编号:4443777 发布者IP:49.5.1.25 浏览:57次![](http://img4.11467.com/u/23565750/194548651.jpg)
- 供应商
- 北京冬璞泰和科技有限责任公司 商铺
- 认证
- 资质核验:已通过营业执照认证入驻顺企:第9年主体名称:北京冬璞泰和科技有限责任公司组织机构代码:911101080649175474
- 报价
- 请来电询价
- 品牌
- 罗氏
- 型号
- 10881767001
- 关键词
- 10881767001
- 所在地
- 北京市海淀区瀚河园131号
- 联系电话
- 010-57181378
- 手机号
- 13716040241
- 联系人
- 马浩然 请说明来自顺企网,优惠更多
产品详细介绍
T7 RNA Polymerase can transcribe RNA from cloned DNA templates
that are downstream from a T7 promoter. The synthesis can be
performed with labeled NTPs to generate highly labeled RNA.
Synthesized RNA can be used in many applications, including:
• RNA or DNA blotting techniques
• In situ hybridization
• RNase protection studies: Transcripts synthesized by the
enzyme are used as precursor RNA for studies on RNA splicing and
processing.
• Synthesis of capped RNA in vitro with addition of
m7GpppG or m7GpppA in excess over GTP or ATP during the
transcription reaction. The generated antisense RNA can be
introduced into cells to suppress the expression of the
corresponding genes.
• Microarray target synthesis
T7 RNA polymerase is commonly used to transcribe DNA which has
been cloned into vectors which have two phage promoters in opposite
orientation. RNA can be selectively synthesized from either strand
of the insert DNA with different polymerases. Homogeneously labeled
single-stranded RNA can be generated with this system. Transcripts
can be non-radioactively labeled with biotin or DIG-11-UTP or
radioactively labeled to high specific activity with
[α-32P] or [α-35S]-labeled nucleotides.
Synthesis of hybridization probes: T7 RNA polymerase allows
highly efficient production of homogeneously labeled RNA. This
labeled RNA may be used as hybridization probes in Southern,
northern, and dot blots, as well as in situ hybridizations.
Suitable labels:Transcripts can be nonradioactively labeled
with biotin-16-UTP or DIG-11-UTP. They may also be radioactively
labeled to high specific activity with [α-32P]- or [α-35S]-labeled
nucleotides.
Test Buffer
40 mM Tris-HCl, pH 8.0 (+20°C), 6 mM MgCl2, 10 mM dithiothreitol, 2
mM spermidine, pH approximately 8.0 (+20°C).
Absence of Endonucleases
1. 1 μg lambda DNA is incubated with T7 RNA polymerase for 4 hours
at +37°C in 25 μl test buffer. The number of enzyme units which
show no degradation of lambda DNA is >100 U.
2. 1 μg Eco RI/Hind III fragments of lambda DNA is incubated with
T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The
number of enzyme units which show no alteration of the banding
pattern is >100 U.
Absence of Nicking Activity
1 μg pBR322 DNA is incubated with T7 RNA polymerase for 4 hours at
+37°C in 25 μl test buffer. The number of enzyme units which show
no relaxing of supercoiled structure is >100 U.
Absence of RNases
4 μg MS2 RNA are incubated with T7 RNA polymerase for 4 hours at
+37°C in 50 μl test buffer. The number of enzyme units which show
no degradation of MS2 RNA is >100 U.
Performance in Transcription Assay
T7 RNA polymerase is function tested in the SP6/T7 Transcription
Kit (Cat. No. 10 999 644 001). The incorporation rate in the
standard assay with 0.5 μg pSPT19 neo DNA linearized with Eco RI
and 50 mCi [alpha-32P] CTP, [400 Ci/mmol (15 TBq/mmol)] gives
>50% of the input radioactivity in 20 minutes.
For life science research only. Not for use in diagnostic
procedures.
Roche has 10x concentrated RNA Labeling Mixes that are specially
designed for DIG- or biotin-labeling. These mixes work well with T7
RNA Polymerase.
1 kit containing 2 components
Preparation NoteActivator: The T7 RNA polymerases are strongly stimulated by BSA or spermidine.
SpecificityPromotor specifity
T7 RNA polymerase is extremely promoter-specific and only
transcribes bacteriophage T7 DNA or DNA cloned downstream of a T7
promoter. Although the T7 and T3 promoter sequences differ only by
3 bp, T7 RNA polymerase only transcribes DNA cloned downstream of
its promoter.
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M
EDTA (pH 8.0) and/or heat to 65 °C.
One unit is the enzyme activity which incorporates 1 nmol CMP in
acid-precipitable RNA products within 60 minutes at +37 °C.
Volume Activity: ≥20 U/μl
Related Categories | |
form | solution |
mfr. no. | Roche |
packaging | pkg of 1,000 U (10881767001) |
pkg of 5,000 U (10881775001) | |
shipped in | dry ice |
storage temp. | −20°C |
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