An highly processive, recombinant (from E.coli strain), thermostable DNA with a very high efficiency of 5’- 3’ polymerase activity and 3´- 5’ exonuclease (non-proofreading) activity. Xtra Taq Pol catalyzes the addition of mononucleotide units to the 3’-end of a primer chain, leading to the formation of DNA products that have 3’-overhanging A nucleotides (thus can be used in TA cloning). This enzyme remains funtional even after prolonged incubation steps at 95°C. The enzyme is supplied at 5U/μl and comes with 5X XtraWhite GL new buffer.

5X XtraWhite GL is a Genespin proprietary formulation, developed for standard and/or high-fidelity amplification of high-GC (>75%) templates. The buffer contains 7.5mM magnesium, PCR enhancers and thickening agents (vortex thoroughly prior to use) and is supplied with 1ml of 6X Orange Loading Dye GL. 5X XtraWhite GL contain an internal fluorescent stain for DNA detection on Agarose gel directly after PCR amplification.

The Orange G dye migrates at the same rate as a duplex DNA fragment of approximately 50 Kbp and does not interfere with DNA migration when it is used as a loading dye for agarose gel electrophoresis.

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Taq Polymerase

FOR RESEARCH USE ONLY

UNIT DEFINITION

One unit is defined as the amount of enzyme required to incorporate 10 nanomoles of dNTPs into acid-insolubile material in 30 min at 74°C.

SHIPPING

Shipped in green ice.

STORAGE

Store at -20C°, store dark.

SHELF LIFE

12 months

FORM

Liquid

CONCENTRATION

5U/ul

component

XSTS-T250 GL

XSTS-Tw250 GL

XSTSn-T250 GL

XSTSn-Tw250 GL

XtraTaq Polymerase

White Buffer

MgCl2

dNTPs

250 units / 50ul

1.5ml 5X XtraWhite Buffer GL

with MgCl2

250 units / 50ul

250 units / 50ul

250 units / 50ul

-

-

100ul / 10mM each

100ul / 10mM each

-

500ul / 50mM

-

500ul / 50mM

component

STS-T1000 GL

STS-Tw1000 GL

STSn-T1000 GL

STSn-Tw1000 GL

XtraTaq Polymerase

White Buffer

MgCl2

dNTPs

1000 units / 200ul

1000 units / 200ul

1000 units / 200ul

1000 units / 200ul

-

500ul / 50mM

-

500ul / 50mM

Assay Set-Up:

Before starting, vortex all components thoroughly to ensure homogeneity.

Prepare a premix for the number of assays you need according to the following protocol:

component

5X Buffer GL

dNTPs

XtraTaq Polymerase

primers

DNA Template

MG Water

stock conc.

5X

10mM each

5U/ul

1ug/ul each

-

-

final conc.

1X

200uM

0.025U/ul

50ng/ul each

-

-

30ul reaction

6.0ul

0.6ul

0.2ul

2.0ul each

10-20ng

up to 30ul

> XtraTaq Pol White GL

Cycling conditions:

Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.

denaturation

denaturation (1)

annealing (2)

extension

95°C

95°C

50-68°C

72°C

5min

30 sec

30 sec

30 sec

4 min

20-35x
1x

1)The annealing temperature depends on the melting temperature of the primers used.

2)The elongation time depends on the length of the fragments to be amplified. A time

of 1 min/kb is recommended.

1.5ml 5X XtraWhite Buffer GL

w/o MgCl2

1.5ml 5X XtraWhite Buffer GL

with MgCl2

1.5ml 5X XtraWhite Buffer GL

w/o MgCl2

4ml 5X XtraWhite Buffer GL

with MgCl2

4ml 5X XtraWhite Buffer GL

w/o MgCl2

4ml 5X XtraWhite Buffer GL

with MgCl2

4ml 5X XtraWhite Buffer GL

w/o MgCl2

-

-

400ul / 10mM each

400ul / 10mM each

GREEN LINE OVERVIEW

5X XtraWhite GL contain an internal fluorescent stain for DNA detection on Agarose gel directly after PCR amplification.

cat. no.

amount

note

XSTS-T5XW 250 GL

XSTS-T5XW 1000 GL

250 units

5X XtraWhite Buffer GL

5X XtraWhite Buffer GL

.PDFMB_end_point_XtraTaq_W_GL_files/StoS%20XtraTaq%20White%20GL%20ds.pdf

XSTS-T5XWw 250 GL

XSTS-T5XWw 1000 GL

1000 units

250 units

5X XtraWhite Buffer  GL w/o MgCl2

5X XtraWhite Buffer  GL w/o MgCl2

cat. no.

amount

note

XSTSn-T5XW 250 GL

XSTSn-T5XW 1000 GL

1000 units

250 units

5X XtraWhite Buffer GL + dNTPs

5X XtraWhite Buffer GL + dNTPs

XSTSn-T5XWw 250 GL

XSTSn-T5XWw 1000 GL

1000 units

250 units

5X XtraWhite Buffer GL w/o MgCl2 + dNTPs

5X XtraWhite Buffer GL w/o MgCl2 + dNTPs

add nucleotides box

1000 units

.PDFMB_end_point_XtraTaq_W_GL_files/StoS%20XtraTaq%20White%20GL%20ds_1.pdf

endpoint PCR

new

green line

*5X XtraWhite Buffer GL is supplied with appropriate quantity of 6X Loading Dye GL

*5X XtraWhite Buffer GL is supplied with appropriate quantity of 6X Loading Dye GL

GeneSpin Srl

Via Friuli, 51

20135 Milano

P.IVA C.F. 04520270960

Laboratory

and sales office

c/o Fondazione Filarete

Viale Ortles, 22/4

20139 Milano

tel. +39 02 56660199

fax. +39 02 537250

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