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HighYield T7 AF488 RNA Labeling Kit

货号 RNT-101-AF488 售价(元) 5475
规格 20T x 20 μl CAS号
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Jena Bioscience

RNT-101-AF488

HighYield T7 AF488 RNA Labeling Kit

20reactions x 20 μl

5475.00

产品简介

HighYield T7 AF488 RNA标记试剂盒设计用于通过体外转录随机产生AF488修饰的RNA探针。这种探针非常适合原位杂交和Northern Blot实验。AF488(也称为Alexa Fluor®488)是一种亲水性染料,与荧光素相比,其光稳定性更强。

UTP-PEG5-AF488使用优化的反应缓冲液和T7 RNA标记聚合酶混合物有效地并入RNA中,作为天然对应物UTP的替代物。35%UTP-PEG5-AF488替代通常会在反应和标记效率之间实现最佳平衡。然而,UTP-PEG5-AF488/UTP比值的单独优化可以通过单核苷酸格式轻松实现。由此产生的AF488修饰RNA探针随后可以通过荧光光谱检测。

该试剂盒含有足够的试剂,用于20个标记反应,每个反应20μl(35%UTP-PEG5-AF488替代物、2.5 mM ATP、GTP、CTP、0.2 mM UTP、0.1 mM UTP-PEG-5-AF488.)。

培养30分钟后,20μl反应产生约5-7μg RNA(1μg T7对照模板,1.4 kb RNA转录物)。然而,产量可能因模板而异(启动子设计、序列长度、二级结构形成)。

Content:
HighYield T7 RNA Labeling Polymerase Mix
2x 40 μl, incl. RNase inhibitor and 50 % glycerol (v/v)

HighYield T7 Reaction Buffer
1x 200 μl (10x), HEPES-based

ATP - Solution
1x 100 μl (100 mM)

GTP - Solution
1x 100 μl (100 mM)

CTP - Solution
1x 100 μl (100 mM)

UTP - Solution
1x 100 μl (100 mM)

UTP-PEG5-AF488
1x 10 μl (5 mM)

T7 G-initiating control template (1.4 kbp)
1x 10 μl (200 ng/μl), 1.4 kbp PCR fragment plus T7 class III phi6.5 promotor resulting in approx. 1400 nt RNA transcript

PCR-grade water
1x 1.2 ml

DTT
1x 100 μl (100 mM)

To be provided by user
T7 Promotor-containing DNA template
RNA purification tools
RNAse-free DNAse I (optional)

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