Primer Design - Design Primer with DNA Sequence (May/28/2009 )
I have been racking my brains. I have a gene of interest: http://www.ncbi.nlm.nih.gov/nuccore/1567852
I need to design primers from a specific list of REs in order to insert the PCR product in to YFP to create a fusion protein.
To do that, I need to determine the promoter region which I don't know.
I have 2 primers from a paper (The Journal of Neuroscience, November 1, 2001, 21(21):8354–8361), used on the cDNA of the above-mentioned gene that were used to insert in to another vector but I cannot use them due to them being non-unique to YFP.
5'-TTAAAATTTGCTAGCACCGCCATGCTGCCCGG-3' (Nhe I)
5-TTTAGCGGCCGCGTTCTGCATCTGCTCAAA-3' (Not I)
Help needed and much appreciated.
I don't know if you want to clone the promoter or cDNA. If you want to clone the cDNA why you want to know the promoter? Here is the refseq for this gene which you should use to design your primer http://www.ncbi.nlm.nih.gov/nuccore/228008403. Most likely you don't need to PCR clone the gene, you can buy the full-length clone from companies at $80 such as http://www.openbiosystems.com/Query/?i=0&q=APP