Scaling down DNA synthesis on ABI 381 A ?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Wed Feb 7 11:56:06 EST 1996


In article <peter.zickert-0702961123220001 at zeke220.onkpat.ki.se>, Peter
Zickert <peter.zickert at onkpat.ki.se> writes
>I woud like to make 100 different 25-mers for RNA flourescent in-situ
>hybridization. 
>
>I have a 381 A with software version 1.23. I wonder if someone have tried
>to scale down the synthesis by diluting the phosphoamidites (about 10
>times) ?
>
>With 40 nmol scale I should get material for 2000 hybridizations ! 200
>would be OK . 
>
>Do I need to upgrade my software to 1.34 or 1.5 ? 
>
>Does anyone have experience with LV columns ?


I have a 381A with 1.23 software. We do 40 nmole synthesis with double
diluted phosphoramidites and modified the cycle program in RAM based on
one from a 391. If I remember correctly the 391 has a couple of
functions that don't exist on the 381 but we just 'ignored' them. I
think the 391 can bubble argon through the column mixing the reagents.
We get roughly 5-7 OD's of oligo for a 22mer. I haven't yet tried the LV
columns. You may have to adjust the Iodine time but that can be done on
the fly.

Regards

Duncan


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