How can I find the sequence after and before my interested ORF? - finding sesequence after and before an ORF? (Dec/22/2005 )
Dear
Hello
I want to clone one of the leishmania gene. But for the first I must design a specific primer and for that I need some sequences that are after and before my favorite gene.
I would try to find them(by NCBI and GENEDB) but I could not find any.
Would you please tell me what can I do? 
What's wrong with using the sequence of the gene itself for your primers?
You do realize that the primers are incorporated into the pcr product (they ARE the PCR product).
-Matt
You do realize that the primers are incorporated into the pcr product (they ARE the PCR product).
-Matt
Thank you for your answering
Really I wanted to design a specific primers. The forward primer have so many similarity to some other sequences in other gene so I decided to find some sequences before that.
Would you please tell me that is it enough to add some sequences regard to the RE to end of my primer in case of using pET system or should I add some sequences regard to His-tag too?
Guilda
You do realize that the primers are incorporated into the pcr product (they ARE the PCR product).
-Matt
Thank you for your answering
Really I wanted to design a specific primers. The forward primer have so many similarity to some other sequences in other gene so I decided to find some sequences before that.
Would you please tell me that is it enough to add some sequences regard to the RE to end of my primer in case of using pET system or should I add some sequences regard to His-tag too?
Guilda
Thank you for your answering
Really I wanted to design a specific primers. The forward primer have so many similarity to some other sequences in other gene so I decided to find some sequences before that.
Would you please tell me that is it enough to add some sequences regard to the RE to end of my primer in case of using pET system or should I add some sequences regard to His-tag too?
Guilda