I want to sequence a specific dicot plant genus with ITS marker for DNA Barcoding test. I've found in some literature that ITS marker is suitable for this specific genus, on which I'm working. Now, I want to know that if I target this ITS region, then is it mandatory to use forward and reverse sequences of both ITS1 and ITS2, or forward primer of ITS1 and reverse primer of ITS2? Please give me suggestions with the primer sequence.
Thanks in advance
Hi! Keya Modak
It is generally advisable to use the primer pair as it is and not interchange (like forward of ITS 1 and reverse of ITS 2).
However, if you really require so, I can suggest you to do an In-silico analysis of the forward primer from ITS 1 together with reverse primer from ITS 2.
If you need any assistance in the analysis, feel free to ask!
Thank you @Arvind Kumar N Sir for your suggestions. Can you send me the pair sequence of the mentioned primer?
Hello! Keya Modak
I suppose you misunderstood my input.
Usually ITS 1 and ITS 2 forward and reverse primers are not interchanged (which means there is ITS 1 forward and reverse primer only used together, and the same for ITS 2).
Since you asked if you can use the forward primer of ITS 1 and reverse of ITS 2 mixed, I suggested you an In-silico (Primer checking) analysis.
Although, if you can mention your plant of interest in personal message, I can provide a previously published ITS 2 primer sequences which can amplify accordingly.
Cheers!
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