SDS-PAGE, AFAIK. Sometimes we use RFLP on PCR products (gain or loss of an enzyme cut site) to confirm a mutation. The polyacrylamide gel - I've never asked, but I've always assumed it gives better separation than agarose.
Here's the really cool thing: we buy premade gels for running digests. Cheers all around!
You do know something: you know how to read all the acronyms I just threw around. :-)
How's your research going, BTW? I haven't seen you post about it recently.
do you do hot RFLPs? they do that here for diagnostics regularly. But not on SDS-PAGE gels, as far as I know. And yeah, makes for a long day for the diagnostic people, from when I've heard them complaining.
Yaaaaay! for premade gels. not that it's difficult to pour a gel, but just annoying.
Well, I kinda felt like I was just pulling things out of nowhere, but I suppose i did know! ;-p
My research is going all right. Tons of work to do, very little time. I've really got to figure something out for after February. My Ph.D. advisor just came to visit and he was very encouraging about my ideas about going back to flies with the mitochondrial neuronal stuff. And he's the head of the Stem Cell Center at Yale now, besides being friendly and well-known in the Drosophila and stem cell fields. Soooooo... I need to think things through and strategize. He figures I need a year to do some pilot drosophila work and get my papers published before starting to apply for faculty type positions. I'm hoping Tom'll be able to get that for me.
I've got some fairly interesting fairly quick experiments going on at the moment and have tons of coverslips that have been stained to analyze, so hopefully the next couple months will get me all I need for at least 2 interesting papers. Current plan is to work all out on collecting tons of data by end of Feb and then write it all up. Should be possible. Just need the cells to work.
I'm in endgame mode now. We'll see if that helps! :-)
Not hot: we PCR a region of interest, digest the product, and run out on polyacrylamide with a ladder. Soak in EtBr and visualize.
Good luck with your research! It sounds like you really do have a lot to do - you're getting near the end of a fellowship, right? I hope some of those coverslips have really awesome data.
Our diagnostics people here put radioactivity into the last cycle of the pcr, and then run it out on the gel. That way the comparisons between the bands can be measured more quantitatively. I think it's also to make faint bands more easily seen.
Yeah, my fellowship ends in February, so busy busy busy.
I just hope the coverslips show what I'm hoping! But I think I'm on a good trajectory now, finally. thanks for the good wishes
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(no subject)
Date: 2008-10-25 03:06 pm (UTC)not PAGE!!! ;-p
Not an SDS-PAGE? Digest, isn't that DNA? Don't you just use agarose for DNA?
Will stop trying to sound like I know something... ;-p
(no subject)
Date: 2008-10-25 06:07 pm (UTC)Here's the really cool thing: we buy premade gels for running digests. Cheers all around!
You do know something: you know how to read all the acronyms I just threw around. :-)
How's your research going, BTW? I haven't seen you post about it recently.
(no subject)
Date: 2008-10-25 07:30 pm (UTC)do you do hot RFLPs? they do that here for diagnostics regularly. But not on SDS-PAGE gels, as far as I know. And yeah, makes for a long day for the diagnostic people, from when I've heard them complaining.
Yaaaaay! for premade gels. not that it's difficult to pour a gel, but just annoying.
Well, I kinda felt like I was just pulling things out of nowhere, but I suppose i did know! ;-p
My research is going all right. Tons of work to do, very little time. I've really got to figure something out for after February. My Ph.D. advisor just came to visit and he was very encouraging about my ideas about going back to flies with the mitochondrial neuronal stuff. And he's the head of the Stem Cell Center at Yale now, besides being friendly and well-known in the Drosophila and stem cell fields. Soooooo... I need to think things through and strategize. He figures I need a year to do some pilot drosophila work and get my papers published before starting to apply for faculty type positions. I'm hoping Tom'll be able to get that for me.
I've got some fairly interesting fairly quick experiments going on at the moment and have tons of coverslips that have been stained to analyze, so hopefully the next couple months will get me all I need for at least 2 interesting papers. Current plan is to work all out on collecting tons of data by end of Feb and then write it all up. Should be possible. Just need the cells to work.
I'm in endgame mode now. We'll see if that helps! :-)
(no subject)
Date: 2008-10-25 08:00 pm (UTC)Good luck with your research! It sounds like you really do have a lot to do - you're getting near the end of a fellowship, right? I hope some of those coverslips have really awesome data.
(no subject)
Date: 2008-10-26 10:23 am (UTC)Our diagnostics people here put radioactivity into the last cycle of the pcr, and then run it out on the gel. That way the comparisons between the bands can be measured more quantitatively. I think it's also to make faint bands more easily seen.
Yeah, my fellowship ends in February, so busy busy busy.
I just hope the coverslips show what I'm hoping! But I think I'm on a good trajectory now, finally. thanks for the good wishes
(no subject)
Date: 2008-10-28 11:59 pm (UTC)