RE: [DIYbio] Gibson assembly master mix

Also, IIRC, chain length is the average length. Maybe it will work but
with low efficiency?

I seldom if ever check molar ratio of backbone to insert for ligation
and never had one fail on me so maybe you can get lucky with small
substitutions? Like Cathal said, its most likely more of an art than a
strict science. On the other hand, at the current street price for
GibbyRxns I would rather play it safe. Do let us know if you attempt it
and if it works. Its nice to add hacks and mods to the list of
protocols.

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D From: Cathal Garvey
Sent: 2/17/2014 5:51 PM
To: diybio@googlegroups.com
Subject: Re: [DIYbio] Gibson assembly master mix
Once you're into long-chain PEG, the difference is often not such a big
deal. Try it with the same weight *or* molarity, ideally both, and see
if it works OK with either.

I'm sure there'll be guides on selecting chain-length for other
protocols, but I wouldn't be surprised if it's more of an art than a
science.

On 17/02/14 08:57, flight505 wrote:
> Hello,
>
> The lab i am working in only has peg 6000, the miller lab protocol
> for Gibson assembly master mix uses peg 8000
> can i adjust for the molecular weight deference and use peg 6000 instead ?
> does anyone have experience with this ?
>
> Best regards,
>

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