d****i
发帖数: 2346 | 1 superfect的protocol还要用PBS洗一遍，发现好多细胞漂起来了，后面再换液一次很多
细胞就被吸走了。 |
s******y
发帖数: 28562 | 2 不要用PBS洗。直接转染即可。293细胞特别容易转染。
如果真的需要洗的话，用含Ca, Mg的PBS
【在 d****i 的大作中提到】
: superfect的protocol还要用PBS洗一遍，发现好多细胞漂起来了，后面再换液一次很多
: 细胞就被吸走了。
|
D******9
发帖数: 2665 | |
d****i
发帖数: 2346 | 4 感谢sunnyday！
今天转染293结果非常惨烈，尽管操作很小心，最后细胞还是所剩无几。 |
m*********7
发帖数: 5207 | 5 Seed 293 cells on ECM coated tissue culture dishes. |
a****d
发帖数: 1919 | 6 293 cells are easily detached from the dish. I normally didn't wash 293
cells if using superfect or Fugene6. Sometimes I used Ca2+ Phosphatase
method and had to wash the cells with PBS, I washed the cells very gently
and quickly.Try not to leave the cells without liquid for too long and add/
remove the PBS very gently. Coating the dish with Gelatin can prevent cell
detaching too. |
d****i
发帖数: 2346 | |
a**v
发帖数: 406 | 8 the easiest (cheapest) way is coating plates with gelatine |
z*y
发帖数: 84 | 9 Does gelatine or poly-lysine affect the normal functions/properties of cells
? |
a*****g
发帖数: 543 | 10 I'm sure 10% FBS and plastic dish affect the normal function/properties of c
ells....
Just teasing....
cells
【在 z*y 的大作中提到】
: Does gelatine or poly-lysine affect the normal functions/properties of cells
: ?
|
|
|
w*****r
发帖数: 2061 | 11 all kinds of artificial crap
c
【在 a*****g 的大作中提到】
: I'm sure 10% FBS and plastic dish affect the normal function/properties of c
: ells....
: Just teasing....
:
: cells
|
a*****g
发帖数: 543 | 12 the harding thing is experimenting with several artificial crap and convinci
ng oneself it not the plastic causing this...
【在 w*****r 的大作中提到】
: all kinds of artificial crap
:
: c
|
c****e
发帖数: 188 | 13 use glycine-coated plate.
when wash cells or change medium, be super gentle.
【在 d****i 的大作中提到】
: superfect的protocol还要用PBS洗一遍，发现好多细胞漂起来了，后面再换液一次很多
: 细胞就被吸走了。
|
t******8
发帖数: 478 | 14 for 293, fugene6 is better. |
z*******a
发帖数: 175 | 15 anybody using Polyethylenimine, such as jetPEI? |
