Re: [DIYbio] Re: Gibson vs SLiCE (single part recommendations)

I haven't tried with the PEG. I recently did another side by side comparison with SLICE made from DH5a cells without pkD46, with pkD46 uninduced and with pkD46 induced. This was just a single shot, so not significant results, but the pkD46 made a big difference, but it didn't matter a much whether it was induced or not. Both SLiCE mixes made with pkD46 gave me about half as many colonies as Gibson Assembly. The reaction was a single gene insert into a plasmid (~1kb and ~5kb) with 25bp overlap on either end.

On Thu, Oct 20, 2016 at 2:36 AM Cathal Garvey <cathal...@cathalgarvey.me> wrote:

Very fair point, yes! This is outside my comfort zone, in terms of calculating the impacts. It's an observation I've made that many protocols allow substitution, but honestly I can't remember whether they specify different molarities.

It would seem to me that yes; longer polymers would mean more soupy buffers. But the difference between PEG50 and PEG3350 is probably orders of magnitude larger than the difference between PEG3350 and PEG8000?

Anyone know good back-of-envelope ways to calculate the relative effects of polymers by chain length? :)

-- @onetruecathal / @cathal@quitter.is

On Wed, Oct 19, 2016 at 7:21 PM, John Griessen <jo...@industromatic.com> wrote:

On 10/19/2016 12:55 PM, Cathal Garvey wrote:

Just an FYI: you can often substitute PEG3350 for PEG8000 in protocols. Just watch for molarity, substituting it gram-for-gram is probably unwise.

Isn't the number after the PEG acronym for length or molecular weight? Its property of interest is absorbing/adsorbing water, so the much bigger molecule at the same number of moles would turn a broth into a pudding. Right?

--

-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@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+un...@googlegroups.com. To post to this group, send email to diy...@googlegroups.com. Visit this group at https://groups.google.com/group/diybio. To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/7f7b1f9e-1cab-b5f5-5b43-25a9d460e1d7%40industromatic.com. For more options, visit https://groups.google.com/d/optout.

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@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+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/1476948978.1963.4%40mail.1984.is.
For more options, visit https://groups.google.com/d/optout.