Hi folks,
crystallized a lot of SRP RNAs - never had success with synthetic RNA,
even with small 30mers T7 RNA pol in vitro transcribed was always
extremely better - moreover: I always use a 3' hammerhead which yields
in the end 100% pure material in mg amounts - Kiyoshi Nagais protocol is
a
board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Martin
Hällberg
Sent: Sunday, March 13, 2011 9:44 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Synthetic RNA for Crystallization
Hi,
I'll second Israels's comment. Since the yield per coupling in synthesis is
lower for RNA than for DNA it gets
Hello All,
I am looking for some advice from some experienced RNA crystallographers. I
would like to order some relatively short (90 bases) synthetic RNAs for
crystallization trials. I was wondering if anyone could comment on the use of
synthetic RNAs for crystallization. Specifically, what is
Hi Michael,
we normally produce synthetic RNAs following this classic paper if the size
is more than let say 40-50 nucleotide, otherwise we buy the RNAs from
Dharmacon and the quality is totally OK.
Hope it helps
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J Mol Biol.
1) Make your own with in vitro transcription (straight T7 r T7+RDRP like in
Finnzymes kit)
2) Buy from IDTDNA - they are very good
Long RNA tends to be expensive. Consider RNA ligase if two or more pieces
can be stitched together.
Artem
On Sun, Mar 13, 2011 at 3:39 AM, Michael Thompson
Hi Mike,
For long RNAs ( 40bases), in vitro transcription is the method of choice.
You might want to take a look at this introductory page from Ambion:
http://www.ambion.com/techlib/basics/transcription/index.html
For structural studies you will have to scale up to milliliter scale. For
that
Hi,
I'll second Israels's comment. Since the yield per coupling in synthesis is
lower for RNA than for DNA it gets really expensive over 30-35 nucleotides.
However, you can stitch together several 30 nt oligos using either T4 RNA
ligase or T4 DNA ligase (with a DNA splint).
Regarding suppliers
