My phosophorylated DNA won't ligate when using the KinaseMax 5'; End-Labeling Kit. What can I do?
We recommend cleaning up the kinase reaction using a phenol/chloroform extraction followed by ethanol precipitation, then eluting the DNA. If this doesn't help, further purify the reaction by spin-column chromatography or denaturing polyacrylamide electrophoresis.
My samples are not labeling well when using the KinaseMax 5' End-Labeling Kit, but the positive control works. What could be the problem?
There may be an inhibitor in the nucleic acid substrate. If needed, clean up the nucleic acid you want to label with a phenol: chloroform: isoamyl alcohol extraction followed by spin-column purification. Alternatively, the 5' end of the RNA may be inaccessible because of tertiary structure. In this case, try heating the RNA to 90°C for 5 minutes, then place it immediately on ice just before the reaction.
What probe kits do you offer for nonisotopic probe labeling and radiolabeled probe labeling?
We offer two nonisotopic probe labeling kits:
- The BrightStar Psoralen-Biotin Nonisotopic Labeling Kit (Cat. No. AM1480)
- MAXIscript Kits for in vitro transcription (Cat. Nos. AM1308 through AM1326)
We offer three radiolabeled probe labeling kits:
- MAXIscript Kits for in vitro transcription (Cat. Nos. AM1308 through AM1326)
- KinaseMax 5' End-Labeling Kit (Cat. No. AM1520)
- DECAprime II DNA Labeling Kit (Cat. No. AM1455)