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Molecular Biology Reagents


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Xtra Taq Pol RTL GL

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 new buffer.

5X XtraRTL 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.

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.

GREEN LINE OVERVIEW

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

cat. no.

amount

note

cat. no.

amount

note

XSTS-T5XRTL 250 GL

XSTS-T5XRTL 1000 GL

250 units

1000 units

5X XtraRTL Buffer GL 

5X XtraRTL Buffer GL

XSTSn-T5XRTL 250 GL

XSTSn-T5XRTL 1000 GL

250 units

1000 units

5X XtraRTL Buffer  GL + dNTPs 

5X XtraRTL Buffer GL + dNTPs 

XSTS-T5XRTLw 250 GL

250 units

5X XtraRTL Buffer GL w/o MgCl2

XSTSn-T5XRTLw 250 GL

250 units

5X XtraRTL Buffer GL w/o MgCl2 + dNTPs 

XSTS-T5XRTLw 1000 GL

PDF

1000 units

5X XtraRTL Buffer GL w/o MgCl2

XSTSn-T5XRTLw 1000 GL

PDF

1000 units

5X XtraRTL Buffer GL w/o MgCl2 + dNTPs 

FOR RESEARCH USE ONLY

SHIPPING

Shipped in green ice.

FORM

Liquid

SHELF LIFE

12 months

STORAGE

Store at -20°C

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.

CONCENTRATION

5U/ul

5X BUFFER

5x conc. complete PCR buffer 

Components

component

XSTS-T5XW 250

STS-T5XWw 250

STSn-T5XW 250

STSn-T5XWw 250

Taq Polymerase

250 units / 50ul

250 units / 50ul

250 units / 50ul

250 units / 50ul

5X RTL  Buffer GL

1.5ml  5X Buffer with MgCl2

1ml 10X Buffer w/o MgCl2

1.5ml  5X Buffer with MgCl2

1.5ml  5X Buffer w/o MgCl2

MgCl2

-

500ul / 50mM

-

500ul / 50mM

dNTPs

-

-

100ul / 10mM each

100ul / 10mM each

component

STS-T5XW 1000

STS-T5XWw 1000

STSn-T5XW 1000

STSn-T5XWw 1000

Taq Polymerase

1000 units / 200ul

1000 units / 200ul

1000 units / 200ul

1000 units / 200ul

5X RTL Buffer GL

4 ml  5X Buffer with MgCl2

4 ml  5X Buffer  w/o MgCl2

4 ml  5X Buffer with MgCl2

4 ml  5X Buffer  w/o MgCl2

MgCl2

-

500ul / 50mM

-

500ul / 50mM

dNTPs

-

-

400ul / 10mM each

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:

component

stock conc.

final conc.

30ul reaction

component

stock conc.

final conc.

30ul reaction

5X RTL Buffer GL*

dNTPs

Taq Polymerase

primers

DNA Template

MG Water

Cycling conditions

5X

10mM each

5U/ul

1ug/ul each

-

-

1X

200uM

0.025U/ul

50ng/ul each

-

-

6.0ul

0.6ul

0.2ul

2.0ul each

10-20ng

up to 30ul

5X Buffer with MgCl2*

5X RTL Buffer GL**

dNTPs

Taq Polymerase

primers

MgCl2

DNA Template

MG Water

5X

10mM each

5U/ul

1ug/ul each

50mM

-

-

1X

200uM

0.025U/ul

50ng/ul each

1.5-4.0 mM

-

-

6.0ul

0.6ul

0.2ul

2.0ul each

0.9-2.4ul 

10-20ng

up to 30ul

5X Buffer without  MgCl2**

Cycling conditions:

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

denaturation

95°C

5min

denaturation (1)

95°C

30 sec

annealing (2)

extension

50-68°C

72°C

30 sec

30 sec

final elong.

72°C

5 min

final step

4°C

forever

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.

Technical Support

Contact us at info@genespin.com to request Technical support with subject and details of your question/inquiry

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