Store at -20℃. For Research Use Only!

dam   ★   65°C

Taq  I

TthHB8 I

Supplied with : 10X Reaction Buffer 'Taq I'

Concentration :            units/㎕

Lot # :                

Expiration date :                

Source : Thermus  aquaticus YT1

Compatible ends (5'…CG)

Aci I, Acl I, Acy I, BmeT110 I, Bpu14 I, Bsa29 I, BsaH I, Bsp119 I, BspAC I, BspD I, BspT104 I, BspX I, BstAC I, BstB I, Bsu15 I, Cla I, Csp45 I, Hap II, Hin1 I, Hin6 I, HinP1 I, Hpa II, Hsp92 I, HpyCH4 Ⅳ, HspA I, Mae II, Mly113 I, Msp I, Nar I, Nsp V, Psp1406 I, Ssi I

Isoschizomers : PpaA II, TthHB8 I

Methylation Sensitivity

: Blocked by overlapping dam methylation

: Not sensitive to dcm and CpG methylation

Unit definition : One Unit is defined as the amount of Taq I required to completely digest 1 ㎍ of λ DNA in one hour at 65℃ in 50 ㎕ assay mixture.

Typical Reaction Conditions : Add 5 ㎕ of 10X Reaction Buffer 'Taq I'[final conc.: 10 mM Tris-HCl (pH 8.4 @ 25℃), 10 mM MgCl₂, 100 mM NaCl, 1 mM DTT], 1~2 ㎕ of DNA(0.5~1.0 ㎍/㎕), 1 ㎕ of Taq I and 42~43 ㎕ of sterile water in 50 ㎕ reaction mixture. Incubate at 65℃.

Heat Inactivation : 80℃ for 20 min.

Storage Conditions : 10 mM Tris-HCl (pH 7.4 @ 25℃), 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 500 ㎍/㎖ BSA, and 50% glycerol. Store at -20℃.

Activity in BeamsBio™ Buffer System

Rxn Buffer	A	B	C	D	'Taq I'
Activity(%)	10~50	10~50	50~75	50~75	75~100

Activity in a complete PCR Mix : 75~100%
Conditions: [10 mM Tris-HCl(pH 8.3 @ 25 ℃), 50 mM KCl, 1.5 mM MgCl₂, 2 pmol of primers, 200 μM dNTP Mix, 2.5 U Taq DNA polymerase and 1 ㎍ of DNA(template or substrate) in 50 ㎕ of reaction volume] After 30 amplication cycles, add 5 units of Taq I. And then incubate at 65℃ for one hour. Relative activity is determined by gel electrophoresis.

No. of Cleavage Sites

λ	Ad2	pUC18	φX174	pBR322	M13mp18	SV40
121	50	4	10	7	13	1

Quality control

Overdigestion: The same band pattern as a digestion in one unit of Taq I for one hour was showing after incubation of 1 ㎍ of λ DNA with         units of Taq I for 16 hr in 50 ㎕ of reaction mixture at 65℃ as determined by agarose gel electrophoresis.

Ligation-Recut: After a 10-fold digestion for one hour,        % of λ DNA fragments were ligated with T4 DNA ligase at 16℃ and        % of the ligated fragments could be recut with Taq I.

Notes

- Conditions of low ionic strength, high enzyme concentration, glycerol > 5%, the presence of Mn²⁺ ion or organic solvents

may result in star activity.

- Taq I is only 0-25% active at 25°C and at 37°C, but retains activity up to 79°C.

References

Sato, S., Hutchison, C.A. III, Harris, J.I. Proc. Natl. Acad. Sci. U. S. A. 74: 542-546 (1977)

Beams Biotechnology Co., Ltd.