DNAzyme-mediated recovery of small recombinant RNAs from a 5S rRNA-derived chimera expressed in Escherichia coli

Table 2 Conditions of 3×pen aRNA cleavage evaluated during optimization of the reaction yield.

Reaction No.	[3×pen aRNA] (μM)*	Molar ratio 3×pen aRNA: Pen17zyme1/1B/1C: Pen17zyme2	Additional components in the annealing buffer	[Mn²⁺] (mM)*	Reaction T (°C)	Time	Additional Cycles
1	0.25	1:0:1	N/A	0	23	10 min	0
2	0.25	1:0:1	N/A	7.5	23	10 min	0
3	0.25	1:0:1	N/A	15	23	10 min	0
4	0.25	1:0:1	N/A	45	23	10 min	0
5	0.25	1:0:1	N/A	15	23	100 min	0
6	0.25	1:1(Pen17zyme1):0	N/A	15	23	100 min	0
7	0.25	1:1(Pen17zyme1B):0	N/A	15	23	100 min	0
8	0.25	1:1(Pen17zyme1C):0	N/A	15	23	100 min	0
9	0.25	1:5(Pen17zyme1B):0	N/A	15	23	100 min	0
10	0.25	1:1(Pen17zyme1B):0	N/A	15	23	17 h	0
11	0.25	1:1(Pen17zyme1B):0	N/A	15	40	100 min	0
12	0.25	1:1(Pen17zyme1B):0	10%EtOH	15	23	100 min	0
13	0.25	1:5(Pen17zyme1B):0	0.3 M KCl, 1.2 M NaCl	15	23	17 h	0
14	0.25	1:5(Pen17zyme1B):0	20% PEG 8000	15	23	17 h	0
15	0.25	1:5(Pen17zyme1B):0	500 μM spermine	15	23	17 h	0
16	0.25	1:5(Pen17zyme1B):0	N/A	15	23	17 h	50
17	0.25	1:5(Pen17zyme1B):0	2.2 M potassium acetate	15	23	17 h	0
18	0.25	1:5(Pen17zyme1B):0	3 M LiCl	15	23	17 h	0
19	0.80	1:1(Pen17zyme1B):1	500 μM spermine	15	23	17 h	0
20	0.80	1:10(Pen17zyme1B):1	500 μM spermine	15	23	17 h	0
21	0.80	1:20(Pen17zyme1B):2	500 μM spermine	15	23	17 h	0
22	0.80	1:10(Pen17zyme1B):10	500 μM spermine	15	23	17 h	0
23	0.80	1:10(Pen17zyme1B):10	500 μM spermine	15	23	17 h	6
24	0.80	1:10(Pen17zyme1B):10	500 μM spermine	15	23	40 h	0
25	0.80	1:10(Pen17zyme1B):10	500 μM spermine	15	23	72 h	0
26	0.80	1:10(bioPen17zyme1B):10(biopen17zyme2)	500 μM spermine	15	23	72 h	0

* The value corresponds to the final concentration of the indicated component in the reaction mixture.

ISSN: 1472-6750