Here is a paragraph in the page of 2014 iGEM Kit:
-- Why Linearized Plasmid Backbones?
Short single stranded DNA fragments will not ligate to 4 bp overhangs. By creating a very short overhang on a PCR of a plasmid backbone, the remnant, when cut with EcoRI and PstI is sufficiently short that it will not anneal at ligation temperature, and will therefore not ligate. This allows us to build high quality construction plasmid backbone without purifying away the cut fragments remaining after PCR.
-----------------------------
And is the linearized one is still a loop-like one? What is the different between the usual one and this one? And how to check if the parts are ligated on the backbone?
Thank you for your help!
-- You received this message because you are subscribed to the Google Groups DIYbio group. 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 https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at http://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/CAPdu10iGiadsarEmyQpY5E1xSpjLv6QBaOs-dSPr0fFBnzrUaw%40mail.gmail.com.
For more options, visit https://groups.google.com/d/optout.
0 comments:
Post a Comment