HELP HELP HELP HELP with pBABE

[mai-mb]

Hi Guys,

I recently switched from a cozy DNA lab into a completely foreign territory of protein based cell culture lab. I came into a lab that has dwindling stalks of its most precious protein and I alone have to find a way to increase and replenish their stalks.

Anyway enough of that.

So they have cloned the 777 sequence (full length) into pBABE. pBabe if I understand correctly from what I have read is a retrovirus?

So my thought processes was that I would take this pBabe grow it up expand it ( is that possible?) and then infect empty phoenix vectors with it. After this is occurred we go ahead an infect the target cells using the Nolen lab protocol.

My Question to you guys is.
Does this make any sense logically.
How do I grow up the pBabe + insert(?) Whats the media whats the protocol?
IS the next step infection of the phenoix cells?
How can I save the grow up pBabe, so I don't lose reserves of that.

Please if anyone could help that would be greatly apprciated, or point in the right direction!!

[mai-mb]

I have also read that the way to get most virus transfected was pBabe(plus insert) + EEV into Bosc23 Cells then use these to target host cells. Anyone know how? Does this mean phoenix cells aren't even a viable choice?