Re: [DIYbio] Re: cheaper/better transformation - using ultrasound (sonoporation)

Success! Here's a picture of pGLO E.coli colonies under UV light, transformed using the $25 35W 42kHz ultrasonic cleaner I bought to clean the bioprinter cartridges.

I haven't seen the exact protocols used for this - and I doubt they did much in terms of optimization for this particular ultrasonic cleaner. But definitely a clear proof of principle...

Patrik


On Tuesday, February 26, 2013 2:59:15 AM UTC-8, Patrik D'haeseleer wrote:
Alright - sounds like we're going to give this a go at BioCurious this week, using the 35W model I got. We'll let you know how it works out...

Patrik

On Monday, February 25, 2013 2:37:30 AM UTC-8, Cathal Garvey wrote:
-----BEGIN PGP SIGNED MESSAGE-----
Hash: SHA256

Does the paper include information on frequency and wattage? I've not
yet tried sonoporation but I'd love to see it tested using
commercially available jewellery cleaners, because there's a lot of
"who knows?" around the issue. If the frequencies are significantly
different, it:
A) Mightn't work at all
B) Might need more or less time to work
C) Might have a different spectrum of useless/useful/murderous activity.

Give it a go! :)

On 02/25/2013 07:27 AM, Patrik D'haeseleer wrote:
>
>
> On Sunday, February 24, 2013 3:56:56 AM UTC-8, Mega wrote:
>
> There once was a sonic cleaner in the store for some 20�.
>
>
> I thought about buying it, but won't it damage the plasmids while
> transforming?????
>
>
> "Ultrasound treatment for 5�60 s results in plasmid transfer to
> /P. putida/ UWC1 (/P/ < 0.05). Exposure time of 10 s gives the
> highest transfer efficiency. No plasmid transfer occurred in the
> absence of ultrasound treatment. (B) Ultrasound reduced /P. putida/
> UWC1 survival following 60 s exposure time, but shorter treatments
> had little effect on bacterial survival (/P/ < 0.05). (C) Plasmid
> DNA was prone to cleavage following ultrasound exposure times
> longer than 30 s"
>
> So yes, you can definitely damage the plasmids and the cells
> themselves with longer exposure times. But you only need a 10s
> exposure to get optimal DNA transfer.
>
> -- -- 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
> http://groups.google.com/group/diybio?hl=en. To view this
> discussion on the web visit
> https://groups.google.com/d/msg/diybio/-/8_-Z_wwWKE4J. For more
> options, visit https://groups.google.com/groups/opt_out.
>
>

- --
Please note my new email: cathal...@cathalgarvey.me
PGP Key: 988B9099
Bitmessage: BM-opSmZfNZHSzGDwdD5KzTnuKbzevSEDNXL
Twitter: @onetruecathal
Code: https://gitorious.org/~cathalgarvey
Blog: http://www.indiebiotech.com
-----BEGIN PGP SIGNATURE-----
Version: GnuPG v1.4.11 (GNU/Linux)
Comment: Using GnuPG with Thunderbird - http://www.enigmail.net/
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=XJ5B
-----END PGP SIGNATURE-----

--
-- 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?hl=en.
To view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/hOm57May0NMJ.
For more options, visit https://groups.google.com/groups/opt_out.
 
 

  • Digg
  • Del.icio.us
  • StumbleUpon
  • Reddit
  • RSS

0 comments:

Post a Comment