Step 1: 50% aqueous acetic acid
Slowly add 100 mL glacial acetic acid to 100 mL distilled water.
Steps 2-5: 1M Triethylammonium bicarbonate, pH 9.4
Dissolve 121 mL triethylamine in 1 L water (final volume)
Bubble CO2 into the solution until pH 9.4
Acidified water, pH 4.6
Bubble CO2 into autoclaved water until pH 4.6
Vydac column buffer A
Component	Final concentration	Stock	Amount
NaH2PO4•H2O	12.5 mM	1 M	12.5 mL
Na2HPO4•7H2O	12.5 mM	0.5 M	25 mL
Adjust to pH 2.8 membrane with glacial acetic acid. Add water to 1 L. Filter through 0.22-μm hydrophilic
Vydac column buffer B
Component	Final concentration	Stock	Amount
NaH2PO4•H2O	62.5 mM	1 M	62.5 mL
Na2HPO4•7H2O	62.5 mM	0.5 M	125 mL
Adjust to pH 2.8 membrane with glacial acetic acid. Add water to 1 L. Filter through 0.22-μm hydrophilic
Nucleoside monophosphate kinase (NM PK) solution
Component	Final concentration	Stock	Amount
NMPK	-	-	60 mg
Tris-HCl, pH 6.5	50 mM	1 M	50 μL
Glycerol	50% (v/v)	100% (v/v)	0.5 mL
Add water to 1 mL
Creatine kinase (CK) solution
Component	Final concentration	Stock	Amount
Creatine kinase	-	-	1 mg
Tris-HCl, pH 7.5	50 mM	1 M	50 μL
Glycerol	50% (v/v)	100% (v/v)	0.5 mL
Add water to 1 mL
Step 6: 10X transcription buffer
Component	Final concentration	Stock	Amount
Tris-HCl, pH 8.0	400 mM	1 M	4 mL
DTT	100 mM	1 M	1 mL
Triton X-100	0.1% (v/v)	10%	0.1 mL
Spermidine, pH 7.0	10 mM	500 mM	0.2 mL
Add water to 10 mL
2X Formamide RNA loading buffer
Component	Final concentration	Stock	Amount
Formamide	95% (v/v)	100% (v/v)	9.5 mL
SDS	0.025% (w/v)	10% (w/v)	25 μL
EDTA, pH 8.0	0.5 mM	50 mM	0.1 mL
Bromophenol blue	0.05% (w/v)	-	5 mg
Xylene cyanol	0.05% (w/v)	-	5 mg
Add water to 10 mL
10X Tris-Borate-EDTA buffer
Component	Final concentration	Stock	Amount
Tris	0.9 M	-	108 g
Boric acid	0.9 M	-	240 g
EDTA, pH 8.0	10 mM	500 mM	20 mL
Add water to 1 L
13% Denaturing polyacrylamide gel (PAGE) solution
Component	Final concentration	Stock	Amount
Acrylamide/bis-acrylamide (19:1)	13%	40%	162.5 mL
Urea	8 M	-	240 g
TBE buffer	1 X	10 X	50 mL
Add water to 500 mL. Protect from light
3 M Sodium acetate, pH 5.2
Dissolve 24.6 g NaOAc in 100 mL water (final volume)