d******r 发帖数: 124 | 1 有什么transfection reagent推荐?我试了磷酸钙,neuroFECT,GenCarrier-2,效果都
不理想,转染效率很低(平均2,3个/8000 neuron),死掉很多。因为需要等轴突树突
分化之后再转染,所以不能用电转和amaxa。用病毒又太费时。各位大侠有什么经验可
以传授吗? |
p*******r 发帖数: 4048 | 2 virus.
【在 d******r 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 有什么transfection reagent推荐?我试了磷酸钙,neuroFECT,GenCarrier-2,效果都 : 不理想,转染效率很低(平均2,3个/8000 neuron),死掉很多。因为需要等轴突树突 : 分化之后再转染,所以不能用电转和amaxa。用病毒又太费时。各位大侠有什么经验可 : 以传授吗?
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d******r 发帖数: 124 | 3 我主要看单个神经元的形态,转染效率并不需要非常高,大概一个波片里有100个转染
成功就足够了。有没有transfection reagent可以达到这种效率?
就是想偷点懒,用病毒工作量太大了:)
【在 p*******r 的大作中提到】![](/moin_static193/solenoid/img/up.png) : virus.
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p*******r 发帖数: 4048 | 4 You can optimize CaPh. It should be able to achive that.
Buy the kit from invitrogen.
【在 d******r 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 我主要看单个神经元的形态,转染效率并不需要非常高,大概一个波片里有100个转染 : 成功就足够了。有没有transfection reagent可以达到这种效率? : 就是想偷点懒,用病毒工作量太大了:)
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d******r 发帖数: 124 | 5 This is what I am doing right now. It's just so frustrating...
I used the kit from Clonetech.
【在 p*******r 的大作中提到】![](/moin_static193/solenoid/img/up.png) : You can optimize CaPh. It should be able to achive that. : Buy the kit from invitrogen.
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p******i 发帖数: 1092 | 6 POLYJET
可以向公司要免费的试用品
http://signagen.com/index.php?main_page=product_info&products_id=657
To request a free trial sample, please Create An Account with us to enter
your shipping address and email us at o***[email protected]
【在 d******r 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 有什么transfection reagent推荐?我试了磷酸钙,neuroFECT,GenCarrier-2,效果都 : 不理想,转染效率很低(平均2,3个/8000 neuron),死掉很多。因为需要等轴突树突 : 分化之后再转染,所以不能用电转和amaxa。用病毒又太费时。各位大侠有什么经验可 : 以传授吗?
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a****d 发帖数: 1919 | 7 for Ca2+ P method, make sure the pH~ 7.04; after adding the mixed plamids,
buffer, wait only half hour, then wash out the transfection medium as clean
as possible(you should be able to see only the black aggregates on the cell
surface, but not other places), then add fresh warm culture medium.
You can also try Fugene HD from Roche, which is better and less toxic
compared to lipofectamin and similar products.
you can also grow cortical neurons on top of glia cells to preserve activity
and help ce |