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

I looked at the specifications for the commercial jewelry sonic cleaners and they are within range of sonoporation protocols -- it's been awhile, but 25 kHz sticks in my head as the spec of typical jewelry cleaners whereas lab equip uses 40 kHz -- well guess what, biology likely doesn't care +/- 50%, it's probable that the lab postdocs authored/optimized the protocol with the equipment they had on hand at the time, which was probably.... 40 kHz!     The freq of the commercial unit will be fixed.  The pressure level at the node is what's important I believe (not really wattage) and might vary quite a bit with cheap units.    But I believe a microbubble agent is required as explained in http://en.wikipedia.org/wiki/Sonoporation  (I'm rather happy of writing that wikipedia entry 'cause it's one of the few I've written that remained factual/undoctored..)

"standard low frequency 40 kHz ultrasound bath" -- doi: 10.1093/nar/gkm710


BTW remind the biohackers not to stick their fingers in the bath...


## Jonathan Cline
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On Monday, February 25, 2013 2:37:30 AM UTC-8, Cathal Garvey wrote:

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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.
>
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