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HighYield T7 AF488 RNA Labeling Kit

貨號(hào) RNT-101-AF488 售價(jià)(元) 5475
規(guī)格 20T x 20 μl CAS號(hào)
  • 產(chǎn)品簡(jiǎn)介
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產(chǎn)品信息

品牌

貨號(hào)

名稱

規(guī)格

單價(jià)

Jena Bioscience

RNT-101-AF488

HighYield T7 AF488 RNA Labeling Kit

20reactions x 20 μl

5475.00

產(chǎn)品簡(jiǎn)介

HighYield T7 AF488 RNA標(biāo)記試劑盒設(shè)計(jì)用于通過(guò)體外轉(zhuǎn)錄隨機(jī)產(chǎn)生AF488修飾的RNA探針。這種探針?lè)浅_m合原位雜交和Northern Blot實(shí)驗(yàn)。AF488(也稱為Alexa Fluor?488)是一種親水性染料,與熒光素相比,其光穩(wěn)定性更強(qiáng)。

UTP-PEG5-AF488使用優(yōu)化的反應(yīng)緩沖液和T7 RNA標(biāo)記聚合酶混合物有效地并入RNA中,作為天然對(duì)應(yīng)物UTP的替代物。35%UTP-PEG5-AF488替代通常會(huì)在反應(yīng)和標(biāo)記效率之間實(shí)現(xiàn)最佳平衡。然而,UTP-PEG5-AF488/UTP比值的單獨(dú)優(yōu)化可以通過(guò)單核苷酸格式輕松實(shí)現(xiàn)。由此產(chǎn)生的AF488修飾RNA探針隨后可以通過(guò)熒光光譜檢測(cè)。

該試劑盒含有足夠的試劑,用于20個(gè)標(biāo)記反應(yīng),每個(gè)反應(yīng)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.)。

培養(yǎng)30分鐘后,20μl反應(yīng)產(chǎn)生約5-7μg RNA(1μg T7對(duì)照模板,1.4 kb RNA轉(zhuǎn)錄物)。然而,產(chǎn)量可能因模板而異(啟動(dòng)子設(shè)計(jì)、序列長(zhǎng)度、二級(jí)結(jié)構(gòu)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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