I was wondering if some of you have experience with pL4440 vector for the in vitro production of dsRNA by using T7 RNA polymerase. How is the vector working when compared with the in vivo protocol inside E.coli?
I did it, worked very well, only it's much more expensive. You get as much as much of T7 and NTPs you want to spend. The cleanup is obviously super easy.
I had troubles with my E. coli strain (they were old and I couldn't determine for sure if they expressed dsRNA), was short on time and had some T7 polymerase that almost expired, so I went for it, but ultimately I hoped to express the dsRNA in vivo to get big amounts without blowing the budget. Unfortunately, that project went on halt, so I can't add more info.
Good luck!
Alex
Alexander... would you please send me a private email to [email protected]. I would like to discuss with you some topics... Thanks
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