Has anyone testet PrimerBank? - (Apr/20/2006 )
I´ve recently come across this page:
Now, I would like to know if someone of you has already sucessfully used their primers and
what your experiences are, like recommendations, "don´t touch it" and so on.
I always design my own. I literally don't trust anyone else to do it. Sorry
me too, John
I rarely rely on someone else's judgement to design a PCR or cloning...just bad juju
i tested two primers for human SLC1A7 gene from primerbank, but they did not work at all. what a pity!
Same for me, they had horrible dimer problems...
When you tested their proposed primers, did you use their protocol or your own and what was your Sequence Detection System (Applied Biosystems?).
Ok. I did it. Ordered 7 primers and tested primer efficiency. Dissociationcurves looked nice for 6 of them, one showed two peaks but for 4 tested samples Cts were above 30 cycles so might be due to small expression. On a gel i got nice bands, no primer dimers.
The RT and real time reactions I did exactly as described in the protocol (http://pga.mgh.harvard.edu/primerbank/PCR_protocol.html). We have the successor of the ABI Prism 7000 and the SDS2.1 Software. Reactionvolume was 10 µl, with 3 µl of a 1/30 dilution for my templates.
I will test another ~10 and if it looks the same I will stick to this software, as it is much faster than designing by myself (yes I´m lazy )
designing your own primers is the only fun part of PCR in my opinion!
well, I usually have to design 40 - 60 primerpairs for one of my projects, and in my opinion to design them all by my own is definitely no fun for me. this time I could use better outdoors, getting a little sun *g* So in the end it all depends, i guess
I have used primer sequences from different primer databases for my housekeeping genes, and they have all worked fine in our lab.