pRF4 selection marker

Model Organism: Worms Methods & Reagents
1

Hi All

I wished to transform pRF4 into bacteria to have a stable copy of this plasmid and wanted to know the selection marker for this rol-6 plasmid.
Thanks

2

It’s amp resistant

3

Yup, it was cloned into pUC19 and is thus AmpR

4

thanks for your replies, could you also tell me what the size of this plasmid is? I have done a miniprep and wished to confirm that I have the correct plasmid.

5

The pRF4 sequence can be found here:
http://forums.wormbase.org/index.php?topic=1522.msg3248

6

An aside: for some applications the rol-6 marker has been superseded. Granted, it is very easy to score using a dissecting scope. But it can cause difficulty later in assessing certain phenotypes. You might want to consider GFP markers in combination with dpy-20 or unc-119 rescue.