艾美捷T7 RNA聚合酶详细说明书

For in vitro use only!

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C

avoid freeze/thaw cycles

Shelf Life: 12 months

Purity: ≥ 95 % (SDS-PAGE)

Form: liquid

Concentration: 2.0 – 2.2 μg/μl (A280, ɛ = 140 L mmol-1 cm-1 [1] in T7 RNA Polymerase HC Storage Buffer)

Description:

Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase (99 kDa) that catalyzes in vitro RNA synthesis from a DNA sequence containing a T7 phage promoter.

The highly concentrated version T7 RNA Polymerase HC is ideally suited for set-up and optimization of high yield in vitro transcription reactions. The provided T7 IVT Set-up Buffer, MgCl2 and DTT solutions enable testing of different reaction conditions such as nucleotide/ MgCl2 concentrations.

Recommendation for assay set-up with minimal optimization: HighYield T7 RNA Synthesis Kit (#RNT-101).

Activity: 1 μl T7 RNA Polymerase HC generates >/= 30 μg RNA in 10 min at 37°. This is comparable to 1000 U of NEB’s T7 RNA Polymerase (High Concentration), #M0460T (activity assay conditions: 1x T7 IVT Set-up Buffer supplemented with 10 mM DTT, 12 mM MgCl2, 3 mM NTPs, 1 μl T7 RNA Polymerase HC and 1 μg DNA template (1.4 kpb RNA transcript) in 20 μl. RNA yield was determined with a fluorescence microplate assay.)

Content:

T7 RNA Polymerase HC

1x 50 μl (2 μg/μl)

50 mM Tris-HCl, 1 mM EDTA, 100 mM NaCl, 5 mM DTT, 0.1 % Triton-X-100, 50 % Glycerol (v/v), pH 8.0

T7 IVT Set-up Buffer

1x 200 μl (10x)

400 mM Tris-HCl (pH 8.0), 20 mM Spermidin

DTT

1x 100 μl (100 mM)

MgCl2

1x 200 μl (200 mM)

Related products:

HighYield T7 RNA Synthesis Kit, #RNT-101

RNAse Inhibitor - recombinant, #PCR-392

NTP bundle, #NU-1014

BIOZ Product Citations:

Please click the arrow on the right to expand the citation list. Click publication title for the full text.

艾美捷T7 RNA聚合酶相关说明：

噬菌体T7 RNA聚合酶是一种DNA依赖性RNA多聚酶（99kDa），对含有T7噬菌体启动子的DNA序列的体外RNA合成进行催化。

高浓缩版T7 RNA聚合酶HC非常适合建立和优化高产量的体外转录反应。所提供的T7 IVT设置缓冲液、MgCl2和DTT溶液能够测试不同的反应条件，例如核苷酸/MgCl2浓度。

具有最小优化的分析设置建议：HighYield T7 RNA合成试剂盒（#RNT-101）。

艾美捷T7 RNA聚合酶相关文献：

1、Novel binding of HuR and poly(C)-binding protein to a conserved UC-rich motif within the 3'-untranslated region of the androgen receptor messenger RNA.

Yeap Bu B; Voon Dominic C; Vivian Julian P; McCulloch Ross K; Thomson Andrew M; Giles Keith M; Czyzyk-Krzeska Maria F; Furneaux Henry; Wilce Matthew C J; Wilce Jackie A; Leedman Peter J;

J Biol Chem (2002) 277:27183-27275

2、Principles of 3' splice site selection and alternative splicing for an unusual group II intron from Bacillus anthracis.

Robart AR, Montgomery NK, Smith KL, Zimmerly S,

RNA (2004) 10:854-862

艾美捷T7 RNA聚合酶组成：

T7 RNA聚合酶HC

1x 50μl（2μg/μl）

50 mM Tris-HCl、1 mM EDTA、100 mM NaCl、5 mM DTT、0.1%Triton-X-100、50%甘油（v/v），pH 8.0

T7 IVT Set-up Buffer

1x 200μl（10倍）

400 mM Tris-HCl（pH 8.0），20 mM精蛋白

DTT

1x 100μl（100 mM）

氯化镁

1x 200μl（200 mM）