产品名称
DIG RNA标记混合物, sufficient for 20 reactions, solution
form
solution
usage
sufficient for 20 reactions
packaging
pkg of 40 μL
manufacturer/tradename
Roche
impurities
Ribonuclease, none detected (up to 20 µl using MSII-RNA)
color
colorless
solubility
water: miscible
storage temp.
−20°C
Quality Level
Analysis Note
经DIG RNA标记试剂盒和DIG核酸检测试剂盒检测的功能。
Application
利用SP6，T7及T3 RNA聚合酶通过体外转录而使用地高辛-11-UTP进行RNA标记。DIG标记RNA已用于多种杂交技术：
- Northern blots
- Southern blots
- 斑点印迹
- 菌斑或克隆提升
- RNase保护实验
- 染色体, 细胞, 及组织切片原位
Biochem/physiol Actions
热灭活：添加 2 μl 0.2 M EDTA (pH 8.0) 终止反应。
Features and Benefits
DIG RNA标记混合物专门设计用于来自Roche并配以优化的转录缓冲液的SP6, T7和T3 RNA聚合酶。
General description
标记效率：来自1μg 线性模板DNA的约10μg的全长地高辛标记RNA被转录。
检测时间：135分钟
样本材料
线性化质粒DNA：
待转录的DNA将会被克隆至含有SP6，T7或T3 RNA聚合酶启动子并相邻于多接头的合适转录载体的多接头位点。对于“流失”转录本的合成，质粒是通过限制性酶进行线性化的。限制性酶生成的5′-突出应当被使用；3′ 突出应当被避免。线性化的模板DNA应当通过酚氯仿抽提及乙醇沉淀进行纯化，以避免RNase的污染。对于′run around′转录环状质粒DNA会被使用。
PCR产物：
含有RNA聚合酶启动子序列的PCR片段也可被用作转录的模板。推荐在转录之前对PCR片段使用HIghPure柱进行纯化。
检测时间：135分钟
样本材料
线性化质粒DNA：
待转录的DNA将会被克隆至含有SP6，T7或T3 RNA聚合酶启动子并相邻于多接头的合适转录载体的多接头位点。对于“流失”转录本的合成，质粒是通过限制性酶进行线性化的。限制性酶生成的5′-突出应当被使用；3′ 突出应当被避免。线性化的模板DNA应当通过酚氯仿抽提及乙醇沉淀进行纯化，以避免RNase的污染。对于′run around′转录环状质粒DNA会被使用。
PCR产物：
含有RNA聚合酶启动子序列的PCR片段也可被用作转录的模板。推荐在转录之前对PCR片段使用HIghPure柱进行纯化。
通过这种方法，特定长度的DIG标记单链RNA探针通过体外转录被生成。在标准条件下，DIG-11-UTP通过SP6、T7和T3 RNA聚合酶以大约每20到25个核苷酸的间隔插入转录产物中。DIG RNA标记混合物专门设计用于优化转录缓冲液的SP6, T7和T3 RNA聚合酶。
便捷的核苷酸混合物用于基于地高辛-11-UTP的RNA标记
内含物
10x 溶液带有：10 mM ATP, CTP, GTP (各), 6.5 mM UTP, 3.5 mM DIG-11-UTP.
便捷的核苷酸混合物用于基于地高辛-11-UTP的RNA标记
内含物
10x 溶液带有：10 mM ATP, CTP, GTP (各), 6.5 mM UTP, 3.5 mM DIG-11-UTP.
Other Notes
仅用于生命科学研究。不可用于诊断。
signalword
Warning
hcodes
Hazard Classifications
Acute Tox. 4 Oral
存储类别
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
does not flash
flash_point_c
does not flash
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商品
Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.
地高辛（DIG）标记方法和试剂盒适用于DNA和RNA DIG探针、随机引物DNA标记、切口平移标记、5'和3'寡核苷酸末端标记。
实验方案
Determine the labeling efficiency in terms of μg (expected yield of a standard labeling reaction is 20 μg of DIG labeled RNA per μg linearized template DNA after the DIG RNA labeling reaction).
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