Date: Thu, 09 Jan 1997 15:14:55 -0700 (MST)
From: "Terry Mulcahy 272-5792; fax 272-9107" <TMULCAHY@medusa.unm.edu>
Subject: DNA Synthesis
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Date: Fri, 13 Sep 1996 14:53:42 -0400
From: "\"\"Dr. Anthony Yeung\"\"" <yeung@deneb.rm.fccc.edu>
Subject: Re: DNAsyn: vials and labels
In-reply-to: "Your message of Fri, 13 Sep 96 10:48:44."
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Hi Dick and Richard:
We can save you some time and money. We make 500 oligos a month. We
elute the oligo off the column on machine with a modified cycle with just 1 ml
of ammonia into a 2 ml vial. The vial fits directly into a Speedvac
polypropylene rotor RH24-18. We dry down the sample in the vial without
needing another evaporator or transfer. If you are concerned about the vacuum
pump, ours has yet to die after 10 years of service this way. We change oil
infrequently, but found that if we keep the pump running except for oil change
and defrosting, nothing will settle to kill the seals. Yes, we are on our 3rd
motor. The pump is vented into the fume hood.
For speedy QC, we have an automated sampler (home made) feeding
samples into a FPLC, running Mono Q anion exchange at pH 12.3. We have
published on that. It is a easy unattended way to know how good your oligos
are by the time you dry them down. Samples are from ammonia after deblocking.
The trityl monitor is good for coupling efficiency, but tell you nothing about
deprotection completeness. The FPLC at pH12.3 has some hydrophobic character
that let you get a feel for the deprotection. Yes, we also use MALDI-TOF,
Perceptive Voyager-DE,sometimes for QC.
Tony
For details, please email Glenn at oligo@fccc.edu
**********************
>Richard,
>
>John Flory at Yale helped us in streamlining our oligo operation and one
>of his best suggestions was to do all manipulations in the 4cc vial that
>screws into the synthesizer. We sent our speedvac 13mm tube rotor to
>Savant along with a 4cc vial with a label on it and they nicely drilled
>out the holes to fit the dna vials.
>
>We deprotect in the vials, put them in a Pierce evaporator to evaporate
>most of the NH4OH and then go directly to the speedvac. They come out of
>the speedvac and go to the researcher. Breakage has never been a
>problem.
>
>The same program we use to track lab work, File Maker Pro, is used to
>print out the labels.
>
>John also suggested All-Pack (Pittsburgh, PA) Teflon caps for the 4cc
>vials which cut the cost per cap from $0.40 to $0.12! They work very
>well.
>
>Regards,
>
>Dick Cook
>MIT
>
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Anthony Yeung, Ph.D. Phone: (215) 728-2488
Member FAX: (215) 728-3647
Fox Chase Cancer Center Internet: AT_Yeung@fccc.edu
7701 Burholme Ave.
Philadelphia. PA 19111
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