Has anyone testet PrimerBank? - (Apr/20/2006 )

Hi,

I´ve recently come across this page:

http://pga.mgh.harvard.edu/primerbank/index.html

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!

http://pga.mgh.harvard.edu/primerbank/index.html

---

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 )

cheers

mike

---

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.

coco

---