In theory the biobrick assembly is a very elegant method, in practice it is often a real pain. Especially when you try to use 3 way ligation for combining a big and a small brick, such as a large protein coding sequence and a small RBS. For the assembly of small bricks I would recommend overlapping PCR instead or when working with multiple parts go for Gibson assembly instead: http://2010.igem.org/Team:Cambridge/Gibson/Introduction
On Saturday, 2 June 2012 17:51:16 UTC+2, Mega wrote:
Once again I came over the term biobrick...--
I know what it should mean (Promotor is one brick, gene another, origin of replication , etc.)
But when you put them together, why are you sure that they will have the right place (1-2-3-4) instead of sticking together anyhow (in a 4-2-1-3 manner) ??
Do they have speciel restriction sites?? That do not attach to itself?
If I understood correctly, one can assemble plasmids (or even entire genoms) using this technique. And if the promoter doesn't work, just take aonther biobrick-promotor. And then it may work.
Maybe a stupid question for one who is familiar with it...
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/dCyRiqSFtwEJ.
To post to this group, send email to diybio@googlegroups.com.
To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com.
For more options, visit this group at http://groups.google.com/group/diybio?hl=en.
0 comments:
Post a Comment