Buffer composition affects ribosome footprint precision in Arabidopsis root and shoot

The goal of this study is to test ionic strength and buffering capacity in polysome extraction buffer on ribosome footprints in Arabidopsis root and shoot. Overall design: Ribo-seq generated from Arabidopsis root and shoot prepared with four different polysome extraction buffers. All four buffers contain 2% polyoxyethylene 10 tridecyl ether, 1% deoxycholic acid, 1 mM DTT, 100 µg/mL cycloheximide, and 10 unit/mL DNase I, but with different ionic strength or buffering capacity listed below. Buffer A: 200 mM Tris-HCl (pH 8), 50 mM KCl, 25 mM MgCl2; buffer B: 200 mM Tris-HCl (pH 8), 40 mM KCl, 20 mM MgCl2; buffer C: 200 mM Tris-HCl (pH 8), 30 mM KCl, 15 mM MgCl2; buffer D: 100 mM Tris-HCl (pH 8), 40 mM KCl, 20 mM MgCl2. 8 libraries in total.