h*********g
发帖数: 18 | 1 I bought a pCMVsport6 vector with one gene inserted. I want to confirm the
sequence of the gene. Using T7 primers, I can get part of the sequence. But
using sp6, M13 forward and reverse primers, I cannot get any sequence. Why?
Can anyone help me about this?
Thanks. |
l******a
发帖数: 3339 | 2 不能用cmv protomoter 来sequence？
But
?
【在 h*********g 的大作中提到】
: I bought a pCMVsport6 vector with one gene inserted. I want to confirm the
: sequence of the gene. Using T7 primers, I can get part of the sequence. But
: using sp6, M13 forward and reverse primers, I cannot get any sequence. Why?
: Can anyone help me about this?
: Thanks.
|
l***y
发帖数: 638 | 3 cmv的sequence google一下就有了，很多测序公司都有cmv的引物
实在不行用你gene的sequence去搞几个primer测几段overlap的自己拼
But
?
【在 h*********g 的大作中提到】
: I bought a pCMVsport6 vector with one gene inserted. I want to confirm the
: sequence of the gene. Using T7 primers, I can get part of the sequence. But
: using sp6, M13 forward and reverse primers, I cannot get any sequence. Why?
: Can anyone help me about this?
: Thanks.
|
h*********g
发帖数: 18 | 4 I have not tried cmv promoter. But sp6 or M13 reverse is more closer to the
3' end of my gene. Any suggestions? Thanks.
【在 l******a 的大作中提到】
: 不能用cmv protomoter 来sequence？
:
: But
: ?
|
h*********g
发帖数: 18 | 5 Next I will use restriction enzyme to cut, and use T7 and SP6 to make probes
. So I want to know the correct sequence. Why sp6, M13forward and reverse
not working? Thanks.
But
?
【在 h*********g 的大作中提到】
: I bought a pCMVsport6 vector with one gene inserted. I want to confirm the
: sequence of the gene. Using T7 primers, I can get part of the sequence. But
: using sp6, M13 forward and reverse primers, I cannot get any sequence. Why?
: Can anyone help me about this?
: Thanks.
|
T*****e
发帖数: 247 | 6 不知道有个小细节楼主注意过没有，有的时候通用引物的序列可能是反的
不过你说完全没有测出来，是引物有问题吗？
如果你有那个插入的gene序列的话，我建议你自己每隔500bp设计一个引物，把这个基
因测通 |
f*****f
发帖数: 195 | 7 买的载体总会有序列，公司的通用引物也该有序列吧。比对一下看看具体在哪有没问题。
-------------------
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是指sequence of insertion还是完全没信号，假如引物跟载体能匹配的话估计就是测
序的问题了，想测哪段自己合成引物也成。 |
