Timeline for Primer design for Gibson assembly
Current License: CC BY-SA 3.0
7 events
| when toggle format | what | by | license | comment | |
|---|---|---|---|---|---|
| Jun 10, 2015 at 22:16 | comment | added | Luigi | This link helps to answer the question, but on Stack Exchange we prefer answers which don't depend entirely on a link to another website | |
| May 27, 2015 at 17:56 | comment | added | Rory M | @MarkB would you be able to edit your post to summarise some of the content of that link in case it is not around in the future? | |
| May 27, 2015 at 17:35 | review | Low quality posts | |||
| May 27, 2015 at 22:56 | |||||
| May 22, 2013 at 12:35 | comment | added | shigeta | Most vectors do have convenient cut sites, but I can see that this is convenient in a lot of ways and it would be quick to order new oligos for the vector. i really want to try this in the lab... | |
| May 22, 2013 at 7:11 | comment | added | Mark B | Well the downside to cutting the vector is that a restriction site must already exist at the junction. This is a major limitation that Gibson overcomes. The potential downsides of using PCR to amplify the backbone: you might have problems getting PCR to work (always a potential problem), and there is a minimal risk of introducing mutations during PCR (but phusion polymerase should give very few mutations). | |
| May 21, 2013 at 12:58 | comment | added | shigeta | it says can linearize the plasmid by cutting it and so you'd only need 2 primers to clone the insert. what advantage is there clone the vector as well? I guess it saves a miniprep, but 2 oligos cost more than a miniprep does.. | |
| May 21, 2013 at 6:03 | history | answered | Mark B | CC BY-SA 3.0 |