MB_GL XtraTaq White

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. 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.

cat. no.

amount

note

PDF

XSTS-T5XW 250 GL

250 units

5X XtraWhite Buffer GL

XSTS-T5XWw 250 GL

XSTS-T5XW 1000 GL

1000 units

250 units

XSTS-T5XWw 1000 GL

1000 units

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

5X XtraWhite Buffer GL

5X XtraWhite Buffer GL w/o MgCl2

add nucleotides box

cat. no.

amount

note

PDF

XSTSn-T5XW 250 GL

250 units

5X XtraWhite Buffer GL + dNTPs

XSTSn-T5XW 1000 GL

1000 units

5X XtraWhite Buffer GL + dNTPs

XSTSn-T5XWw 250 GL

XSTSn-T5XWw 1000 GL

250 units

1000 units

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

5X XtraWhite Buffer GL w/o MgCl2 + dNTPs

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°

SHELF LIFE

12 months

FORM

Liquid

CONCENTRATION

5U/ul

GREEN LINE OVERVIEW

5x White Buffer GL contain an internal fluorescent stain for DNA detection on Agarose gel directly after PCR amplification.

component

XSTS-T5XW 250 GL

XSTS-T5XWw 250 GL

XSTSn-T5XW 250 GL

XSTSn-T5XWw 250 GL

XtraTaq Polymerase

250 units / 50ul

White Buffer

1.5ml 5X XtraWhite Buffer

GL with MgCl2

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

MgCl2

-

500ul / 50mM

-

500ul / 50mM

dNTPs

-

100ul / 10mM each

component

XtraTaq Polymerase

XSTS-T5XW 1000 GL

1000 units / 200ul

XSTS-T5XWw 1000 GL

XSTSn-T5XW 1000 GL

1000 units / 200ul

XSTSn-T5XWw 1000 GL

1000 units / 200ul

White Buffer

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

MgCl2

-

500ul / 50mM

-

500ul / 50mM

dNTPs

-

400ul / 10mM each

component

5X Buffer GL

dNTPs

XtraTaq Polymerase

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:

stock conc.

final conc.

5X

1X

10mM each

200uM

5U/ul

0.025U/ul

6.0ul

0.6ul

0.2ul

30ul reaction

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

50-68°C

72°C

5min

30 sec

4 min

primers

DNA Template

1ug/ul each

-

50ng/ul each

-

2.0ul each

10-20ng

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.

MG Water

-

up to 30ul

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