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| Helps reduce non-specific signals. If I remember correctly, the random primers sometimes generate much smaller labelled fragments (that is whey they are random) which can give you a lot of non-specific bands in your northern blot. I used to use some sort of spin columns which did not take more than 10 minutes of my time. It helps a lot. Best of luck Jay -----Original Message----- From: [Only registered users see links. ] [mailto:[Only registered users see links. ].indiana.edu] On Behalf Of Simone Marker Sent: Monday, August 27, 2007 9:06 AM To: [Only registered users see links. ] Subject: purification of radioactive probe Hi, does anyone know if it is really necessary to purify a radioactive DNA probe after labelling it by random priming? I want to use it on a northern blot. So far, I worked with DIG-labelled probes, that do not need to be purified after labelling. They are used directly in the hybridisation solution. Is it different with radioactive probes? Do you think that precipitation by salt and alcohol is sufficient for purification? Thank you very much, Simone _______________________________________________ Methods mailing list [Only registered users see links. ] [Only registered users see links. ] This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. |
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| probe , purification , radioactive |
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| Thread | Thread Starter | Forum | Replies | Last Post |
| Real Time-PCR probe design | Ganto | Real-Time PCR and Quantitative PCR Forum | 1 | 03-22-2010 08:00 PM |
| minimum radioactive probe size | Ed Siefker | Protocols and Methods Forum | 1 | 09-26-2007 08:50 PM |
| purification of radioactive probe | Simone Marker | Protocols and Methods Forum | 2 | 08-28-2007 12:13 PM |
| Re:purification of radioactive probe | Analia Alet | Protocols and Methods Forum | 0 | 08-27-2007 11:14 PM |
| problems with probe binding to 28S and 18S RNA only on Northerns | Wendy Ingram | Protocols and Methods Forum | 0 | 09-11-2006 02:30 AM |