Hi,

I don't know which part of the plant you are using but perhaps you should consider mature rosette rather than young seedling and to wound the leaves before extracting if you can't or don't want to use an elicitor.

Dear Megha,

There is report stating that in Arabidopsis microRNA miR172b regulates the transcription of FLS2. The level of miR172b is very low in the early stage of seedling development but increases over time, which results in increased FLS2 transcription. Hence try using cDNA from at least one week old plants.

Reference: doi: 10.1105/tpc.18.00297

Best Regards,

Kishor

Dear Megha,

You should try after infection because basal level of FLS2 is very low and it induces after infection.So you can give a try after infection.

Hi Kishor,

I have clone AtFLS2 CDS in pDONR221, if needed I can share an aliquot with you.

Best,

Caterina

Thanks to all of you for sharing your suggestions. I was using the leaf tissue from 1-2 weeks old Col-0 plants. I will use different combinations this time (Age and/or infection). Thanks to all of you.