AB Bioscience LLC
"The Home of KlenTaq1"
 Formerly AB Peptides since 1992
Woman Owned Small Business
Authorized Vendor to US Gov

GF Klentaq1 Cat# 1003



(A Glycerol-Free version of Klentaq1 DNA Polymerase)

Amount: 100 µl (sufficient for 2000 25 µl reactions up to 1 kb)

Shipping conditions: Ice pack

Storage conditions: Enzyme at 2-8°C, buffer -20°C

*Note: Do not freeze enzyme.

Shelf life: Not determined. (Suggesting the enzyme to be used soon after delivery.)

 

PRODUCT DESCRIPTION:

Klentaq1 is a 5'-exonuclease deficient Taq polymerase (an N-terminal deletion of Taq) with improved fidelity and thermostability. 10x buffer composition is: 500 mM Tris-Cl pH 9.2, 160 mM ammonium sulfate, 0.5% Brij 58, and 35 mM magnesium chloride. We also offer (upon request) 10x buffer at pH 7.9 for better fidelity.





TYPICAL PCR PROTOCOL for a 25 ml reaction:

Reagent

Volume

Final Concentration

10x Klentaq1 Reaction buffer

2.5 ml

1x

dNTP mix (10 mM)

0.5 ml

200 mM each

Left Primer

variable

0.2 mM

Right Primer

variable

0.2 mM

DNA template

variable

0.1-100 ng

 

 

 

Klentaq1 Polymerase**

0.05 ml

 

de-ionized distilled H2O

Adjust final volume to 25 ml

-

DNA amount depends mostly on genome size and target gene copy number.

**To determine specific optimal enzyme concentration, we strongly recommend an enzyme titration test for each target. Targets larger than 1 kb may require more enzyme or may benefit from the use of an LA (Long Accurate) version of the polymerase.

 

 

CYCLING CONDITIONS

1. Denaturing:                      94° for 2 minutes for 1 cycle

2. Denaturing:                     94° for 20-30 seconds

3. Annealing:                        50°-68° depending on the specific Tm primers for 40-60 seconds

4. Extension:                        68° for 2 min / 1kb target

5. Repeat steps 2-4 for 25-40 cycles

 

 

 

 

REFERENCES:

Barnes, W.M. (1994) PCR amplification of up to 35 kb DNA with high fidelity and high yield from bacteriophage templates, PNAS 91, 2216-2220.

 

U.S. Patent No. 5,436,149