Re: [DIYbio] How do you store your DNA constructs?

[Jeswin]

On Sun, Feb 10, 2013 at 6:45 PM, Edward Perello <edw...@deskgen.com> wrote:
> How is everyone freezing and archiving their DNA? This includes for those of
> you working at labs - what is the rule of thumb, how do you order it, do you
> keep spreadsheets of it all? How much is there? This is for plasmids,
> oligos, primers etc
>

Well, all our DNA is in -20 freezer. The oligos are dissolved in TE
and stored in boxes with capacity for probably 81 tubes. I got a
spreadsheet with location and name of oligo. Plasmids are stored in TE
or other elution buffer, H2O. For low copy, I use TE+BSA(1:100). I
don't track plasmids in Excel. Just take from the appropriate box as
needed.

Primers are oligos, right? Or am I missing something?

[Nathan McCorkle]

On Feb 12, 2013 2:33 PM, "Jeswin" <phill...@gmail.com> wrote:

> Primers are oligos, right? Or am I missing something?
>

Yes but oligos can be used as primers or other

[Dakota Hamill]

Yeah right now I have DNA(genomic) and primers stored in H2O.  Should probably think about switching to TE but have been using them for PCR and then sending them for sequencing and they recommend to avoid TE due to Mg2++ chelation.

But...the first 2 sequences that we had run turned out crap (extremely low confidence), didn't even pass the quality assurance test.  They sat over the weekend (4 days) so I assume nucleases chewed them apart...So maybe TE would have been better after all!

[Eric Ma]

Online lab catalog with one labmate shared on it in case something happens to me.

• Primers sequentially numbered with sequence & description
• Plasmids sequentially numbered with description, specification on antibiotic resistance and origin of copy # (for quick reference)

Primer stocks: 100 µM in IDTE buffer, stored in -20C; 10 µM working stocks diluted with sterile distilled water, room temperature. I have had primers stored at RT for over 3-4 months now, and they still work.

Plasmids: -20C until I need them; they can stay on my bench for a few days max before I put them back, but sometimes I get a bit sloppy and they're on my bench for ~2 weeks.

I specifically do not use TE buffer for miniprepped plasmids, because I need to co-transform them with other plasmids in my biological machine, and the salts sometimes cause arcing. 

Cheers,
Eric
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On Sun, Feb 10, 2013 at 6:45 PM, Edward Perello <edw...@deskgen.com> wrote:

How is everyone freezing and archiving their DNA? This includes for those of you working at labs - what is the rule of thumb, how do you order it, do you keep spreadsheets of it all? How much is there? This is for plasmids, oligos, primers etc

Cheers!

Ed

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[Mega]

What I do to store plasmids at home... 

Take a marmelade jar (empty & clean of course). Put in about one centimeter of water. Freeze it. 

Then put the plasmids in. 

Because if the freezer does unfreeze (to get the ice away from it's pipes) cycle, the water acts as a cold sink and keeps the air and the plasmid cold... 

(I don't even know if my freezer can defrost, because it is quite old. But better on the safe side.