Purification, reconstitution, and mass analysis of archaeal RNase P, a multi-subunit ribonucleoprotein enzyme

. Author manuscript; available in PMC: 2022 Sep 25.

Published in final edited form as: Methods Enzymol. 2021 Sep 25;659:71–103. doi: 10.1016/bs.mie.2021.07.006

Component	Initial concentration	Volume (μL)	Final concentration
T7 RNAP IVT buffer	10X	10	1X
DTT	100 mM	10	10 mM
NTP mix	25 mM each	20	5 mM each
TIPP	0.2 U/μL	0.5	0.001 U/μL
T7 RNAP IVT template	See note 12
T7 RNAP	See note 5
ddH₂O	Adjust reaction to final volume of 100 μL