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
cat. no.
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 XtraWhite 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.
XSTS-T5XW 250
250 units
5X XtraWhite Buffer
XSTS-T5XW 1000
1000 units
5X XtraWhite Buffer
XSTS-T5XWw 250
250 units
5X XtraWhite Buffer w/o MgCl2
XSTS-T5XWw 1000
1000 units
5X XtraWhite Buffer w/o MgCl2
*5X XtraWhite Buffer is supplied with appropriate quantity of 6X Loading Dye
add nucleotides box
XSTSn-T5XW 250
250 units
5X XtraWhite Buffer + dNTPs
XSTSn-T5XW 1000
1000 units
5X XtraWhite Buffer + dNTPs
XSTSn-T5XWw 250
250 units
5X XtraWhite Buffer w/o MgCl2 + dNTPs
XSTSn-T5XWw 1000
1000 units
5X XtraWhite Buffer w/o MgCl2 + dNTPs
*5X XtraWhite Buffer is supplied with appropriate quantity of 6X Loading Dye
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
component
XSTS-T5XW 250
XSTS-T5XWw 250
XSTSn-T5XW 250
XSTSn-T5XWw 250
XtraTaq Polymerase
250 units / 50ul
250 units / 50ul
250 units / 50ul
250 units / 50ul
White Buffer
1.5ml 5X XtraWhite Buffer
with MgCl2
1.5ml 5X XtraWhite Buffer
w/o MgCl2
1.5ml 5X XtraWhite Buffer
with MgCl2
1.5ml 5X XtraWhite Buffer
w/o MgCl2
MgCl2
-
500ul / 50mM
-
500ul / 50mM
dNTPs
-
-
100ul / 10mM each
100ul / 10mM each
component
XSTS-T5XW 1000
XSTS-T5XWw 1000
XSTSn-T5XW 1000
XSTSn-T5XWw 1000
XtraTaq Polymerase
1000 units / 200ul
1000 units / 200ul
1000 units / 200ul
1000 units / 200ul
White Buffer
4ml 5X XtraWhite Buffer
with MgCl2
4ml 5X XtraWhite Buffer
w/o MgCl2
4ml 5X XtraWhite Buffer
with MgCl2
4ml 5X XtraWhite Buffer
w/o MgCl2
MgCl2
-
500ul / 50mM
-
500ul / 50mM
dNTPs
-
-
400ul / 10mM each
400ul / 10mM each
component
5X Buffer
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
95°C
50-68°C
72°C
5min
30 sec
30 sec
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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