An hack to SLiCE that I found very useful in the lab...
https://wiki.london.hackspace.org.uk/view/SPLiCE
Basically you add like %3 PEG 8k to the reaction mixture and boom it works much better.
I have used SLiCE and SPLiCE for assemblies up to 7 parts and around 2kb long for gene synthesis and mostly codon optimizing genes for expression in other organisms. The most important part is the overlap regions in my experience, where they should be basically unique with respect to one another.
2017 Motohashi protocol on Springer is overall a good starting point on optimizing your reaction. - http://link.springer.com/protocol/10.1007%2F978-1-4939-6472-7_23
Good luck!
Cihan Aydin, Ph.D. Linkedin: www.linkedin.com/in/cihanaydinphd RG: www.researchgate.net/profile/Cihan_Aydin
On 04/16/2017 01:55, Rob C wrote:
Good question, I haven't worked with repeats with slice.--
I suspect it'll be heavily dependent on repeat size, number of repeats, content, linkers, where the repeats are in the plasmid relative to place of intended insertion and homology ends, etc. Hard to speculate really...only one way to know for sure. ;)
On Saturday, April 15, 2017 at 3:40:04 AM UTC-7, Mega [Andreas Stuermer] wrote:What if your plasmid contains repeats? Will there be unwanted recombination?
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