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how to chose right subsequent for her2(erbb2)gene ?


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#1 Bassam27

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Posted 02 April 2012 - 02:07 PM

I NEED URGENT HELP
How to chose right sequences for her2(erbb2)gene to design primer ? i want to detect the level of over expression of gene ?

I choose accession number (NM_004448) for HER2 (ERBB2) this is variant 1

Transcript Variant: This variant (1) represents the shorter


transcript but encodes the longer isoform (a) .

And there is variant 2 where t

his variant (2) contains additional exons at


its 5' end and lacks an alternate 5' noncoding exon, compared to
variant 1. These differences result in translation initiation at an
in-frame, downstream AUG and an isoform (B) with a shorter
N-terminus compared to isoform a
.
i do not know what is differents between them ???????????? plz help me

#2 Trof

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Posted 03 April 2012 - 10:06 AM

You need to find out which variant you are looking for, which one is significant for your aplication.
If you're not sure, design primers that detect both.

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#3 michaeltaozmkm

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Posted 30 March 2014 - 01:34 AM

The universal primer is:

 

5' sequencing primer: CMV-F
3' sequencing primer: BGHrev


#4 bob1

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Excellent

Posted 30 March 2014 - 01:39 PM

 

The universal primer is:

 

5' sequencing primer: CMV-F
3' sequencing primer: BGHrev

 

Only for particular plasmids - this won't work for the OP's original question as they don't have a cloned gene - they are amplifying from RNA/cDNA.






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