Hypervariable V4 region of 16S rRNA (E. coli positions 515-806).
Even "perfect" in silico designs should be verified before ordering: Primer3 Input (version 0.4.0) primer3 0.4.0
Ensuring primers bind to DNA at the exact right temperature [10]. GC Content: Aiming for 40-60% to ensure stability [11]. Secondary Structures: Hypervariable V4 region of 16S rRNA (E