cat. no.
amount
note
cat. no.
amount
note
250 units
1000 units
250 units
1000 units
5X XtraWhite Buffer GL
5X XtraWhite Buffer GL
5X XtraWhite Buffer GL w/o MgCl2
5X XtraWhite Buffer GL w/o MgCl2
250 units
1000 units
250 units
1000 units
5X XtraWhite Buffer GL + dNTPs
5X XtraWhite Buffer GL + dNTPs
5X XtraWhite Buffer GL w/o MgCl2 + dNTPs
5X XtraWhite 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 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 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 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 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.
