g*********r
发帖数: 59 | 1 I wonder whether anyone has experience on this retrvirus vector. Tried to
amplify it several time. the plasmid yield from most clones are super low.
Any way to solve this issue?
THanks! |
a*****n
发帖数: 2835 | 2 try to transform again to DH5a,and make new preparations.
【在 g*********r 的大作中提到】
: I wonder whether anyone has experience on this retrvirus vector. Tried to
: amplify it several time. the plasmid yield from most clones are super low.
: Any way to solve this issue?
: THanks!
|
F**********e
发帖数: 158 | 3 you are right, this vector get a very low yeild
transform again may help a little
we just use much more bacteria |
s******y
发帖数: 28562 | 4 Yes, it is very low.
What I did is I picked up several colonies from the plate and then compare
their plasmid yields.
The highest one will be used for plasmid production.
Even so, I still have to shake 500mL culture in order to get 250ug plasmids
【在 g*********r 的大作中提到】
: I wonder whether anyone has experience on this retrvirus vector. Tried to
: amplify it several time. the plasmid yield from most clones are super low.
: Any way to solve this issue?
: THanks!
|
g*********r
发帖数: 59 | 5 Glad to hear that. I thought I might make some mistakes during midi prep.
I also notice that sequence provided by addgene is not right. I tried to cut
it with kpni.the bands I got differs from predict. |
X***n
发帖数: 366 | 6 DH5a or Invitrogen Stbl3? It says Stbl3 is better for retroviral vector.
【在 a*****n 的大作中提到】
: try to transform again to DH5a,and make new preparations.
|
