16s rRNA seq of the maize leaf microbiome
1-cm wide strips of leaf tissue were cut from the margin to the midrib halfway between tip and auricle. Tissue from 3 plants was bulked, frozen in liquid nitrogen, and stored at -80C. Total RNA was extracted with a Qiagen Rneasy 96 kit and the V5/V6 region of 16s rRNA reverse-transcribed with primer 1391R (GACGGGCGGTGWGTRCA) then amplified with primers 799F/1115R (Shade et al 2013; doi: 10.1128/mBio.00602-12)