BAC DNA purification

Use this category for questions directly related to DNA manipulation (isolation, purification, sequencing, etc.) and questions regarding general PCR methodologies

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Re: BAC DNA purification

Postby 29yrsExperience » Aug 01 2018 6:28 pm

Understood. I guess I am running out of ideas. You can try the bacterial transformation test to see if it does appear to be BAC DNA you've got, but I don't know if that means it is pure enough for microinjection. Also, looking at the DNA on a pulsed-field gel apparatus, if there is one available in your institution, might give you better imagery. Good luck!
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Re: BAC DNA purification

Postby kiruphagaran » Aug 01 2018 6:59 pm

Thanks for the suggestions. I will try transformation and I'm looking for pulse-field apparatus in our university.
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Re: BAC DNA purification

Postby r.rosati » Aug 02 2018 4:48 pm

kiruphagaran wrote:Hi all,
This is regarding the BAC prep in my previous post. I am purifying 106 kb BAC DNA. I get 2 forms of BAC DNA ( I don't know whether it is supercoiled and circular single stranded or nicked plasmid). But iam unable to attach the gel pic here. Could someone help with it?

Thanks
kirupa


29yrsExperience wrote:Hi Kirupa,

I have not been able to upload a photo to this site in a while; tried yesterday, in fact. I suspect something is a little wrong with the website. For a few weeks I could not even log in.
So, does anyone else know how to get the photo uploading function fixed? I know a lot about cloning, but diddley about websites.


Hi,
thank you for your comments, apparently there is indeed an issue with image uploading to the site.
I'm just a voluntary moderator, but I know for a fact that the inability to log in was due to the migration of the site to a different server - that's also why we went from "molecularbiology.forums.biotechniques.com" to "forums.biotechniques.com".
I will notify the site admins about this, I'm sure they'll start looking into it right away. In the meanwhile, however, I would suggest uploading the picture to a third-party free image-hosting website (such as imgur.com) and copy the link here.
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