Hi all,
I am planning to over-express a lncRNA (under 35S) in Arabidopsis. Will the presence of 20-30 bp sequences upstream and downstream of lncRNA sequence affect its over-expression? I understand that for mRNA cloning, cloning primers should must include some bp at the 5' end.
Any suggestions or comments are welcome.
Thanks much.
Uwe Borgmeyer
You have to choose a vector with a suitable promoter and a terminator. When you generate your PCR product covering the full-length cDNA, all you have to add, are 6 bp on each site (restriction enzyme sites). Each primer should include a few extra base pairs for sufficient cutting at their 5'-ends, however these will be lost during cloning.
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