Merck - 11175025910 - DIG RNA Labeling Kit (SP6/T7)

ITEM #: 13106563
Storage: -20C
UNSPSC Code: 41106000
General description: Sigma Life Science is committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical.-á The DIG System was established as a sensitive and cost-effective alternative to using radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.
General description: Assay Time: 145minutes. Sample Materials. Linearized plasmid DNA. PCR product Principle. The DIG RNA Labeling Kit produces DIG-labeled, single-stranded RNA probes of known length. Either SP6 or T7 RNA polymerase transcribes these probes in vitro from template DNA (in the presence of digo x igenin-UTP).RNA Labeling by in vitro Transcription. The DNA to be transcribed is cloned into the polylinker site of appropriate transcription vectors (e.g., pSPT 18 or 19), which contain promoters for SP6 and T7 RNA polymerases. Adjacent template DNA is linearized at a suitable site and the RNA polymerases are used to produce "run off" transcripts. DIG-UTP is incorporated into the transcript. Every 20 to 25th nucleotide of the newly synthesized RNA is a DIG-UTP. Since the nucleotide concentration does not become limiting in the standard transcription reaction, this reaction can generate large amounts of labeled RNA.
General description: Kit for the labeling of RNA with digo x igenin-UTP by in vitro transcription with SP6 and T7 polymerases. By this method, single-stranded RNA probes of known length are produced, which can be used in a variety of hybridization techniques.
Application: For RNA labeling with DIG-11-UTP by in vitro transcription with SP6 and T7 RNA Polymerases. DIG-labeled "run off" transcripts are synthesized with high efficiency and can be used in a variety of hybridization techniques: . Northern blots. Southern blots. In situ hybridizations. Plaque or colony lifts. RNase protection e x periments. Due to highly specific and sensitive detection systems, DIG-labeled probes can be used for single-copy gene detection in 1++g total human : Since the linkage between DIG and UTP is resistant to alkali, DIG-labeled RNA can be fragmented by alkaline treatment. Slightly reducing the size of the DIG-labeled RNA probe may make it more suitable for certain applications in in situ hybridization.
Quality: Test principle: The DNA template to be transcribed is cloned into the polylinker site of appropriate transcription vectors, which contain promoters for SP6 and/or T7 RNA Polymerases. After linearization at a suitable site, RNA is transcribed in the presence of DIG-UTP. Under standard conditions, appro x imately 10mug of full-length DIG-labeled RNA is transcribed from 1mug template.
Specificity: Sensitivity and SpecificityDIG-labeled RNA probes can detect single-copy genes in as little as 1 mug of mammalian DNA under the following assay conditions: The hybridization mi x contains 20 to 100 ng labeled probe/ ml, and the bound probe is detected with anti-DIG-AP and visualized with the chemiluminescent substrate CDP- inactivation: Stop the reaction by adding 2 mul 0.2 M EDTA (pH 8.0).
Other Notes: For life science research only. Not for use in diagnostic procedures.
DIG RNA Labeling Kit (SP6/T7)
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