RNA Polymerase SP6
更新时间:2016-12-01 10:48:04 信息编号:4443798 发布者IP:49.5.1.25 浏览:65次![](http://img4.11467.com/u/23565750/194549095.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
- 10810274001
- 关键词
- 10810274001
- 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
Application
SP6 RNA Polymerase can transcribe RNA from cloned DNA templates
that are downstream from an SP6 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.
With this system, homogeneously labeled single-stranded RNA can
be generated. 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.
Contents:
• SP6 RNA Polymerase, ≥20 U/μl, in buffer, pH 7.9
• Transcription Buffer, 10x concentrated
仅用于生命科学研究。不可用于诊断。
Preparation NoteActivator: BSA/spermine
QualityTest 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 SP6 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 > 30 U.
2. 1 μg Eco RI/Hind III fragments of lambda DNA is incubated with
SP6 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 > 30 U.
Absence of Nicking Activity
1 μg pBR322 DNA is incubated with SP6 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 > 30.
Absence of RNases
4 μg MS2 RNA are incubated with SP6 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 > 30.
Performance in Transcription Assay
SP6 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 pST18 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.
Promoter specifity
SP6 RNA polymerase is extremely promoter-specific and only
transcribes bacteriophage SP6 DNA or DNA cloned downstream of a SP6
promoter (e.g., pSPTBM20; pSPTBM21).
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
Synthesis of hybridization probes: SP6 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, DIG-11-UTP, or fluorescein-12-UTP. They may
also be radioactively labeled to high specific activity with
[a-32P]- or [a-35S]-labeled nucleotides.
Note: Roche has 10x concentrated RNA Labeling Mixes that are
specially designed for DIG-, biotin-, and fluorescein-labeling.
These mixes work well with SP6 RNA Polymerase.
Related Categories | |
form | solution |
mfr. no. | Roche |
packaging | pkg of 1,000 U (10810274001) |
pkg of 5,000 U (11487671001) | |
shipped in | dry ice |
storage temp. | −20°C |
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