100 mm mgso4

UltraPure™ Tris-HCI pH 8.0, RNase free EDTA, RNase free MgCl₂, RNase free KCl, Novex™ TBE Running Buffer (5X), 2X TBE-Urea Sample Buffer, Novex™ TBE-Urea Gels, 15%, SYBR^® Gold Nucleic Acid Gel Stain, and SYBR^® Green II RNA Gel Stain were purchased from Thermo Fisher Scientific (Waltham, MA). Nuclease-free water and oligo length standard 10/60 were purchased from Integrated DNA Technologies, Inc. (Coralville, IA). Nt.BstNBI nicking endonuclease, Bst 2.0 WarmStart^® DNA Polymerase, 10× ThermoPol I Buffer, dNTPs, BSA, and 100 mM MgSO₄ were purchased from New England Biolabs (Beverly, MA).
Oligonucleotides were ordered from two different sources to avoid trigger contamination in templates. Desalted amplification templates were purchased from Integrated DNA Technologies (Coralville, IA) suspended in IDTE Buffer at a concentration of 100 µM. Templates were modified with an amino group on the 3’ end to prevent template extension. All desalted trigger oligonucleotides were purchased from Eurofins Genomics (Louisville, KY) suspended at a concentration of 50 µM in TE Buffer. Triggers were diluted in Nuclease-free water in a separate room to prevent contamination.