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 XtraRTL (Ready To Load) new buffer.

5X XtraRTL Reaction Buffer is a Genespin proprietary formulation, developed for standard and/or high-fidelity amplification of high-GC (>75%) templates. The buffer contain 7.5mM magnesium, PCR enhancers and thickening agents (vortex thoroughly prior to use).

5X XtraRTL Reaction Buffer contains Orange G dye that allows gel loading and electrophoresis of the sample directly from the PCR tube, without further manipulation. 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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You are here: molecular biology / endpoint PCR / XtraTaq Pol RTL

endpoint PCR

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°

SHELF LIFE

12 months

FORM

Liquid, orange

CONCENTRATION

5U/ul

component

XSTS-T5XRTL 250

XSTS-T5XRTLw 250

XSTSn-T5XRTL 250

XSTSn-T5XRTLw 250

XtraTaq Polymerase

RTL Buffer

MgCl2

dNTPs

250 units / 50ul

1.5ml 5X XtraRTL Buffer

with MgCl2

250 units / 50ul

250 units / 50ul

250 units / 50ul

-

-

100ul / 10mM each

100ul / 10mM each

-

500ul / 50mM

-

500ul / 50mM

component

XSTS-T5XRTL 1000

XSTS-T5XRTLw 1000

XSTSn-T5XRTL 1000

XSTSn-T5XRTLw 1000

XtraTaq Polymerase

RTL 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

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 RTL (ready to load)

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 XtraRTL Buffer

w/o MgCl2

1.5ml 5X XtraRTL Buffer

with MgCl2

1.5ml 5X XtraRTL Buffer

w/o MgCl2

4ml 5X XtraRTL Buffer

with MgCl2

4ml 5X XtraRTL Buffer

w/o MgCl2

4ml 5X XtraRTL Buffer

with MgCl2

4ml 5X XtraRTL Buffer

w/o MgCl2

-

-

400ul / 10mM each

400ul / 10mM each

cat. no.

amount

note

XSTS-T5XRTL 250

XSTS-T5XRTL 1000

250 units

5X XtraRTL Buffer

5X XtraRTL Buffer

.PDFMB_end_point_XtraTaq_RTL_files/StoS%20XtraTaq%20Pol%20RTL%20%20ds.pdf

XSTS-T5XRTLw 250

XSTS-T5XRTLw 1000

1000 units

250 units

5X XtraRTL Buffer w/o MgCl2

5X XtraRTL Buffer w/o MgCl2

cat. no.

amount

note

XSTSn-T5XRTL 250

XSTSn-T5XRTL 1000

1000 units

250 units

5X XtraRTL Buffer + dNTPs

5X XtraRTL Buffer + dNTPs

XSTSn-T5XRTLw 250

XSTSn-T5XRTLw 1000

1000 units

250 units

5X XtraRTL Buffer w/o MgCl2 + dNTPs

5X XtraRTL Buffer w/o MgCl2 + dNTPs

add nucleotides box

1000 units

.PDFMB_end_point_XtraTaq_RTL_files/StoS%20XtraTaq%20Pol%20RTL%20%20ds_1.pdf

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20135 Milano

P.IVA C.F. 04520270960

Laboratory

and sales office

c/o Fondazione Filarete

Viale Ortles, 22/4

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tel. +39 02 56660199

fax. +39 02 537250

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