n*j
2 楼
狗的爸今天问了一堆业余问题,什么通知没收到怎么办,真服了 。。。
S*e
3 楼
我自己也还在干活,养了一大堆细胞等着做knockdown。我总是遇到一个问题,当我准
备transfect 293 细胞的时候,他们都一片一片的掉下来了, 这样plasmid还能进去吗
?大家怎么解决这个问题的。
Happy Holiday!
备transfect 293 细胞的时候,他们都一片一片的掉下来了, 这样plasmid还能进去吗
?大家怎么解决这个问题的。
Happy Holiday!
c*d
4 楼
祝水过
n*k
6 楼
either too confluent, or you didn't handle with care...or your cells have
been too old etc...Make sure you cells
are fine and then handle with care, never overgrow them...Alternative, coat
plates with poly-Lysine, still never
overgrow the cells...
【在 S**********e 的大作中提到】
: 我自己也还在干活,养了一大堆细胞等着做knockdown。我总是遇到一个问题,当我准
: 备transfect 293 细胞的时候,他们都一片一片的掉下来了, 这样plasmid还能进去吗
: ?大家怎么解决这个问题的。
: Happy Holiday!
been too old etc...Make sure you cells
are fine and then handle with care, never overgrow them...Alternative, coat
plates with poly-Lysine, still never
overgrow the cells...
【在 S**********e 的大作中提到】
: 我自己也还在干活,养了一大堆细胞等着做knockdown。我总是遇到一个问题,当我准
: 备transfect 293 细胞的时候,他们都一片一片的掉下来了, 这样plasmid还能进去吗
: ?大家怎么解决这个问题的。
: Happy Holiday!
b*e
7 楼
bless
l*e
9 楼
应该你操作不小心,细胞可能状态不好, 不过 和上面说的想反, 如果你细胞过少的
话, 也容易脱。
话, 也容易脱。
R*0
10 楼
祝顺利通过!
R*0
13 楼
过了吗?求签经
a*d
32 楼
I coated the plates with gelatin/PBS to prevent cells detaching. If you
handle the cells carefully enough, you don't really have to coat the plates.
When you changing medium, do one plate per time, and don't try to change
medium for more than 3 plates in one shot. Cells don't like being exposed to
air for too long. Another thing is to, add medium or PBS to the wall but
not directly to the cells.
handle the cells carefully enough, you don't really have to coat the plates.
When you changing medium, do one plate per time, and don't try to change
medium for more than 3 plates in one shot. Cells don't like being exposed to
air for too long. Another thing is to, add medium or PBS to the wall but
not directly to the cells.
l*e
35 楼
antibiotics does not matter if you use fugene6. I remember the company state
it in the protocol.
it in the protocol.
Z*5
37 楼
养了一年半细胞了,从来没有用过抗生素,也没有污染过。
另外我转染都用罗氏的FuGene HD,不用换液,操作简单。 我们实验室转293T的都用这
个。
另外我转染都用罗氏的FuGene HD,不用换液,操作简单。 我们实验室转293T的都用这
个。
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