m******5
发帖数: 1383 | 1 在agrose胶上，RNA product看起来多大？
我在胶上跑出来都是和我的DNA templete同样大小的东西，几乎重合在一起，似乎不大
make sense? |
z*******6
发帖数: 679 | |
a****o
发帖数: 1786 | 3 to check RNA, you should run a denature gel, either agarose+MOPS or
polyacralimide+urea. If your RNA intends to form no 2nd structure, it may
run at similar position as its DNA template. otherwise something is wrong.
Single strand DNA or RNA gives very low signal compared to the same size ds
DNA if you stain with EtBr. |
m*****n
发帖数: 760 | 4 for fast examination, you could run your RNA samples on 2% TBE agrose gel.
for accurancy, you want to run them on denaturing PAGE gel.
【在 m******5 的大作中提到】
: 在agrose胶上，RNA product看起来多大？
: 我在胶上跑出来都是和我的DNA templete同样大小的东西，几乎重合在一起，似乎不大
: make sense?
|
m*****n
发帖数: 760 | 5 if you are more comfortable with the old-school RNA gel,
check details at http://www.ambion.com/techlib/append/supp/rna_gel.html
I have used this before, and it worked very well.
good luck.
【在 m******5 的大作中提到】
: 在agrose胶上，RNA product看起来多大？
: 我在胶上跑出来都是和我的DNA templete同样大小的东西，几乎重合在一起，似乎不大
: make sense?
|
m*****n
发帖数: 760 | 6 if you are more comfortable with the old school agrose gel,
check for details at http://www.ambion.com/techlib/append/supp/rna_gel.html
I have used this protocol before, and it worked very well.
good luck.
【在 m******5 的大作中提到】
: 在agrose胶上，RNA product看起来多大？
: 我在胶上跑出来都是和我的DNA templete同样大小的东西，几乎重合在一起，似乎不大
: make sense?
|
m******5
发帖数: 1383 | 7 I am still wondering why it runs around the similar position as its DNA
template. RNA product is ssRNA while the DNA templete is dsDNA…………
ds
【在 a****o 的大作中提到】
: to check RNA, you should run a denature gel, either agarose+MOPS or
: polyacralimide+urea. If your RNA intends to form no 2nd structure, it may
: run at similar position as its DNA template. otherwise something is wrong.
: Single strand DNA or RNA gives very low signal compared to the same size ds
: DNA if you stain with EtBr.
|
m*********n
发帖数: 215 | 8 Your RNA forms secondary structure. that's why the guy upstairs told you to
run it on a DENATURE gel to kill the secondary structure.....
【在 m******5 的大作中提到】
: I am still wondering why it runs around the similar position as its DNA
: template. RNA product is ssRNA while the DNA templete is dsDNA…………
:
: ds
|
f*******e
发帖数: 628 | 9 实在是要跑一般的胶的话，要在 formamide 里面 heat denature，然后snap cool 到
冰上，马上跑胶。不过里面 DNA 会出 double band。 |
