Dear ccp4
I am working on hbx (Hepatitis B x protein for both that duck and the human)
I worked first on the dhbx which contained the vector pRSET-A and I didn’t get
any results, then I changed the vector ( thanks to many of you who provided me
with information, papers and books) using p-MAL c2x which worked perfectly
throughout the whole procedure, I even now have about 60% soluble protein, then
I started working on the hhbx,also changing the vector from pRSET-A to p-MAL
c2xat first the PCR was successful with an obvious band on the gel, so I
proceeded with double digestion using both restriction enzymes which are BamH1
and Hind111, incubated 3h then applied the insert on the gel, surprisingly what
I observed was a slight band on the gel, after purifying the insert I checked
the concentration which decreased from 35ng/ul before purification to 5.2ng/ul
after purification, I would be grateful if any one has any idea or suggestions
regarding this situation
Best Regards
Rana
