cDNA kit recommendations?
Dr. Duncan Clark
news at hgmp.mrc.ac.uk
Sat Mar 24 05:59:42 EST 2001
In article <99ggj0$nnb$2 at testinfo.cs.uoguelph.ca>, the eminent Allison
MacKay at University of Guelph wrote
>I need to make a cDNA library from a novel plant species
>(i.e. NO pre-made libraries exist, alas!). I have not in
>fact ever actually made a library before and I was wondering
>if people had recommendations for which kits to use/not use?
>I am particularly interested in any brands which I should
>avoid, as I'd rather not deal with poor product in addition
>to my own inexperience. Any advice is appreciated!
A few things you will need to decide:
What vector are you going to make the library in - lambda phage or a
plasmid. The choice here could influence primers for the cDNA synthesis.
Also expression library or not.
You will need a proper cDNA synthesis kit not a more common first strand
You will need to prepare high quality RNA, as I would think you want as
long a length clones as possible. This could be prepared by a kit or a
home made solution.
Are you going to select for poly-A tailed RNAs? I would have thought yes
so then you need to consider what sort of oligodT purification column,
filters etc. to use.
I would do a test run on your final choice of kits before you go near
them with valuable RNA sample!
Stratagene always used to be one of the companies catalogues to look at
first because they had a good range of lambda cDNA synthesis
kits/vectors. Amersham used to be reasonable for I haven't checked there
catalogues for these kits for a while. Have a look at Invitrogen/LTI and
possibly Clontech. My guess would be a simple choice between Invitrogen
and Stratagene if you want an all in one kit.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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