Long-Range PCR Amplification of DNA by DNA Polymerase III Holoenzyme from Thermus thermophilus

Table 3

Summary of additives tested to increase the thermostability of Tth pol III holoenzyme.

Cosolvents	Sugars	Crowding agents	Detergents
Glycerol (5–25%)^a; Sorbitol (5–25%); Mannitol (2.5–10%); Maltitol (5–20%); 1-Methyl-pyrrolidinone; (5–20%); 1-Methylindole (5–20%); 2-Pyrrolidinone (5–20%); Acetamide (0.25–1 M); Trimethylamine N-oxide; (10 mM–1 M); Tertiary butane (5–20%); Trimethyl ammonium; chloride (50 mM–1 M); Methylsulfonylmethane; (0.12–6%)	Trehalose; (140–720 mM); Sucrose; (60 mM–1.2 M); β-Cyclodextrin; (0.72–7.2 mM); α-Cyclodextrin; (0.84–8.4 mM); Glucose (3–21%); D-Fructose (5–20%); D-Mannose (5–20%); D-Galactose (2–20%); Arabinose (5–80 mM)	PEG 400 (1–5%); PEG 4000 (2–5%); PEG 8000 (2–5%); PEG 20000 (2–5%); CM cellulose; (0.16–1.2%); Polyvinylpyrrolidone; (0.01–3%); Polyvinyl alcohol; (0.5–4%); Ficol (0.5–3%)	Tween 20^b; NP-40; Pluronic acid; Zwittergent 3-08; Zwittergent 3-10; Zwittergent 3-12; Zwittergent 3-14; Zwittergent 3-16; Chaps; Chaps SO; N-Octyl-sucrose; Caprolyl Sulfobetaine; Myristyl-sulfobetaine; SB 3-10; SB 3-14; N-Octyl-β-glucopyranoside; N-Octyl-β-D-thioglucopyranoside
Betaines	Salts	Metals	Other
NDSB 195 (50 mM–1 M); NDSB 201 (50 mM–1 M); NDSB 256 (0.5 mM–1 M); 3-1-Pyridino-1-propan-sulfonate (50 mM–1 M); Betaine monohydrate; (0.25–2 M); Betaine hydrochloride; (0.25–1.25 M)	Potassium glutamate; (25–200 mM); Sodium acetate; (25–200 mM); Sodium citrate; (25–200 mM)	Zinc sulfate; (0.25–2 μM); Magnesium sulfate; (0.5–4 μM)	L-Proline (0.12–1.2 M); Ethylene glycol tetraacetic acid (EGTA) (5–25 mM)

Values in parenthesis represent ranges of concentrations tested.
^bDetergents were tested at two concentrations, at CMC and at 5X lower than CMC.