|
DIG RNA Labeling Kit (SP6/T7)
For RNA labeling with digoxigenin-UTP by in vitro transcription with SP6 and T7 RNA polymerase.
Application
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 northern blots, Southern blots, in situ hybridization, plaque or colony lifts, and RNase protection experiments.
Contents
1.pSPT18 DNA, 40 µl, 0.25 mg/ml
2.pSPT19 DNA, 40 µl, 0.25 mg/ml
3.Control DNA 1, pSPT18-Neo, 20 µl, 0.25 mg/ml cleaved with Pvu II
4.Control DNA 2, pSPT19-Neo, 20 µl, 0.25 mg/ml cleaved with Pvu II
5.DIG-labeled Control RNA, 100 µl, 100 ng/µl DIG-labeled “antisense” neo RNA dimethyldicarbonate treated double-distilled water
6.Unlabeled Control RNA, 20 µl, 200 µg/ml neo poly (A) “sense” RNA in dimethyldicarbonate treated double-distilled water
7.NTP Labeling Mixture, 40 µl, 10x conc. 10 mM ATP, CTP, GTP, 6.5 mM UTP, 3.5 mM DIG-11-UTP, pH 7.5
8.Transcription Buffer, 40 µl, 10x conc.
9.DNase I, RNase-free, 40 µl, 10 U/µl
10.Protector RNase Inhibitor, 20 µl, 20 U/µl
11.SP6 RNA Polymerase, 20 µl, 20 U/µl
12.T7 RNA Polymerase, 20 µl, 20 U/µl
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, approximately 10 µg of full-length DIG-labeled RNA is transcribed from 1 µg template.
|