MB_GL XtraTaq RTL

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 GL new buffer.

5X XtraRTL 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 XtraRTL 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

XSTS-T5XRTL 250 GL

5X XtraRTL Buffer GL

250 units

XSTS-T5XRTL 1000 GL

1000 units

5X XtraRTL Buffer GL

XSTS-T5XRTLw 250 GL

5X XtraRTL Buffer GL w/o MgCl2

250 units

XSTS-T5XRTLw 1000 GL

1000 units

5X XtraRTL Buffer GL w/o MgCl2

add nucleotides box

cat. no.

amount

note

XSTSn-T5XRTL 250 GL

250 units

5X XtraRTL Buffer GL + dNTPs

XSTSn-T5XRTL 1000 GL

1000 units

5X XtraRTL Buffer GL + dNTPs

XSTSn-T5XRTLw 250 GL

250 units

5X XtraRTL Buffer GL w/o MgCl2 + dNTPs

XSTSn-T5XRTLw 1000 GL

1000 units

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

component

XSTS-T5XRTL 250 GL

XSTS-T5XRTLw 250 GL

XSTSn-T5XRTL 250 GL

XSTSn-T5XRTLw 250 GL

XtraTaq Polymerase

250 units / 50ul

RTL Buffer

1.5ml 5X XtraRTL Buffer

GL with MgCl2

1.5ml 5X XtraRTL Buffer

GL w/o MgCl2

1.5ml 5X XtraRTL Buffer

GL with MgCl2

1.5ml 5X XtraRTL Buffer

GL w/o MgCl2

-

500ul / 50mM

-

500ul / 50mM

MgCl2

dNTPs

-

100ul / 10mM each

component

XSTS-T5XRTL 1000

XSTS-T5XRTLw 1000 GL

XSTSn-T5XRTL 1000 GL

XSTSn-T5XRTLw 1000 GL

XtraTaq Polymerase

1000 units / 200ul

RTL Buffer

4ml 5X XtraRTL Buffer

GL with MgCl2

4ml 5X XtraRTL Buffer

GL w/o MgCl2

4ml 5X XtraRTL Buffer

GL with MgCl2

4ml 5X XtraRTL Buffer

GL w/o MgCl2

-

500ul / 50mM

-

500ul / 50mM

MgCl2

dNTPs

-

400ul / 10mM each

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:

Cycling conditions:

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

denaturation

95°C

5min

component

final conc.

stock conc.

30ul reaction

denaturation (1)

95°C

30 sec

annealing (2)

50-68°C

30 sec

5X Buffer GL

5X

1X

6.0ul

dNTPs

200uM

0.6ul

10mM each

extension

72°C

30 sec

4 min

XtraTaq Polymerase

5U/ul

0.025U/ul

0.2ul

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.

primers

1ug/ul each

50ng/ul each

2.0ul each

DNA Template

-

10-20ng

MG Water

-

up to 30ul