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TaqI
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TaqI Technical Brochure Technical Brochure

TaqI is a restriction enzyme purified from Thermus aquaticus YT I.

 

Unit substrate: Lambda DNA (dam-).

 

Unit calculation assay conditions: 100 mM KCl, 20 mM Tris-HCl (pH 8.5 @ 25°C), 3 mM MgCl2, 0.04% Triton X-100, 100 μg/ml BSA. Incubate at 65oC.

 

Absence of contaminants: 100 units of TaqI do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of λ DNA (dam-) at 65oC. After 50-fold overdigestion with TaqI, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

 

Storage buffer: 300 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml BSA and 50% glycerol. Store at -20oC

 

Heat inactivation: 80oC for 20 minutes.

 

Methylation Sensitivity:

dam methylation: Blocked by overlapping

dcm methylation: Not sensitive

CpG methylation: Not sensitive

 

Note: Incubation without BSA results in 50% activity. 

 

Percent Activity in MINOTECH Buffers
 L M  H  SH  A  K 
 10-25 50-75  75-100  50-75  50  50 
 
 
General reaction mixture:
10U TaqI 1μl
10x UTaqI buffer*  2μl 
DNA substrate  <1μg
Sterile ultrapure water  Up to 20 μl
Incubate for 15 min at 65oC  
*We recommend the addition of BSA to a final concentration of 100 μg/ml. 

 

Reagents supplied: 10x UTaqI buffer