m*p
发帖数: 226 | 1 Embryonic hearts were co-transfected by RFP and GFP plasmids. Only 20% were
transfected. Boss argued that
all RFP and GFP should co-localized and I said it is impossible, only when
RFP and GFP plasmids are in the
same lipid vesicle and fused with the same cell, then they will colocalize.
She said she was one thousand
percent sure. I can not say more, as this might harm the relationship.
However, I would like to know the truth about this co-transfection. Thanks
for your imput. | a***e
发帖数: 1010 | 2 if you check under microscope, you will find your boss is right. | m*p
发帖数: 226 | 3 As I saw more Red cells than Green cells and some percentage overlap, so my
boss argued something wrong.
Why you think RED AND GREEN colocalize? Thanks! | t******y
发帖数: 716 | 4 if you mixed well, you should be able to see what your boss expected. The
reason is simple that both plasmids have the same chance binding to every
single lipofectamine 2000 particle(if you're using it as transfection
reagent).Each lipofectamine 2000 particle probably can bind a bunch of DNA
and sometimes the particle size is pretty big,ranged from ~40 nm to ~180 nm
in diameter. There is a paper talking about DNA and transfection particles.
http://www.biomedcentral.com/1472-6750/8/23 | s******y
发帖数: 28562 | 5 这个完全取决于转染的效率，其实并不一定的。
反正我做的时候，一般只有50~80%的细胞是两个荧光都有的。
were
.
【在 m*p 的大作中提到】
: Embryonic hearts were co-transfected by RFP and GFP plasmids. Only 20% were
: transfected. Boss argued that
: all RFP and GFP should co-localized and I said it is impossible, only when
: RFP and GFP plasmids are in the
: same lipid vesicle and fused with the same cell, then they will colocalize.
: She said she was one thousand
: percent sure. I can not say more, as this might harm the relationship.
: However, I would like to know the truth about this co-transfection. Thanks
: for your imput.
| l*****a
发帖数: 1431 | |
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