2 replies to this topic

[akhilesh004]

Hay,

I am not getting my beta-actin pcr as insilico pcr and wet lab pcr beta actin length is different.
If someone can help me with this problem, I would be really thankful. If I have missed out some information, please feel free to ask.
Thanks in advance.

[baccilus]

akhilesh004, on 13 September 2010 - 02:58 AM, said:

Hay,

I am not getting my beta-actin pcr as insilico pcr and wet lab pcr beta actin length is different.
If someone can help me with this problem, I would be really thankful. If I have missed out some information, please feel free to ask.
Thanks in advance.

You mean to say that the primer you designed using primer3 like softwares is giving you an amplicon size which is different from the theoretical amplicon size?

[akhilesh004]

baccilus, on 13 September 2010 - 03:16 AM, said:

akhilesh004, on 13 September 2010 - 02:58 AM, said:

Hay,

I am not getting my beta-actin pcr as insilico pcr and wet lab pcr beta actin length is different.
If someone can help me with this problem, I would be really thankful. If I have missed out some information, please feel free to ask.
Thanks in advance.

You mean to say that the primer you designed using primer3 like softwares is giving you an amplicon size which is different from the theoretical amplicon size?

Yes, the band of primer3 is 336 bp and i am getting 250 bp in wet lab PCR amplification.