http://www.stevecheckley.co.uk/blog/2010/06/29/project-planning/ a mind forever voyaging through the strange seas of thought Fri, 03 Feb 2012 13:42:41 +0000 hourly 1 http://wordpress.org/?v=3.3.1 http://www.stevecheckley.co.uk/blog/2010/06/29/project-planning/comment-page-1/#comment-596 Steve Fri, 09 Jul 2010 09:37:54 +0000 http://www.stevecheckley.co.uk/blog/?p=264#comment-596 I've had a look around openwetware.org. I also find the forums on bitesize bio can be useful. My cloning problem was my boss wanting to do directional cloning using a SalI and XhoI combination and refusing to entertain the acknowledged problems of using this combination. I had major problems as my insert was a PCR product which new england biolabs has acknowledged SalI has a problem with. I ended up starting again with different restriction sites and just ignoring the boss. The recent round of cloning worked 1st time (surprise surprise). I will remember to have a route around on openwetware in future though as it can be useful to compare a number of protocols before starting. I haven't encountered ligation by selection and will investigate this one. :) I’ve had a look around openwetware.org. I also find the forums on bitesize bio can be useful. My cloning problem was my boss wanting to do directional cloning using a SalI and XhoI combination and refusing to entertain the acknowledged problems of using this combination. I had major problems as my insert was a PCR product which new england biolabs has acknowledged SalI has a problem with. I ended up starting again with different restriction sites and just ignoring the boss. The recent round of cloning worked 1st time (surprise surprise). I will remember to have a route around on openwetware in future though as it can be useful to compare a number of protocols before starting. I haven’t encountered ligation by selection and will investigate this one.

]]> http://www.stevecheckley.co.uk/blog/2010/06/29/project-planning/comment-page-1/#comment-593 t Fri, 09 Jul 2010 07:34:12 +0000 http://www.stevecheckley.co.uk/blog/?p=264#comment-593 Check out openwetware.org they have a lot of good protocols. If you don't mind sharing, what are the cloning steps that you are getting bogged down in? In my experience, things can be sorted out rather quickly if you have the right setup (GFP dropout, proper flanking restriction sites, use of ligation-by-selection) Check out openwetware.org they have a lot of good protocols. If you don’t mind sharing, what are the cloning steps that you are getting bogged down in? In my experience, things can be sorted out rather quickly if you have the right setup (GFP dropout, proper flanking restriction sites, use of ligation-by-selection)

]]>