cat. no.
amount
note
cat. no.
amount
note
STS-P 250
STS-P 1000
250 units
1000 units
5X XtraWhite Buffer
5X XtraWhite Buffer
STSn-P 250
STSn-P 1000
250 units
1000 units
5X XtraWhite Buffer + dNTPs
5X XtraWhite Buffer + dNTPs
STS-Pw 250
250 units
5X XtraWhite Buffer w/o MgCl2
STSn-Pw 250
250 units
5X XtraWhite Buffer w/o MgCl2 + dNTPs
STS-Pw 1000
1000 units
5X XtraWhite Buffer w/o MgCl2
STSn-Pw 1000
1000 units
5X XtraWhite Buffer 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
2U/ul
5X BUFFER
5 x conc. complete PCR buffer
Components
component
STS-P250
STS-Pw250
STSn-P250
STSn-Pw250
Pfu Polymerase
250 units / 125ul
250 units / 125ul
250 units / 125ul
250 units / 125ul
5X Buffer
1.5ml 5X Buffer with MgCl2
1.5ml 5X 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-P1000
STS-Pw1000
STSn-P1000
STSn-Pw1000
Pfu Polymerase
1000 units / 500ul
1000 units / 500ul
1000 units / 500ul
1000 units / 500ul
5X Buffer
4 ml 5X Buffer with MgCl2
4 ml 5X Buffer w/o MgCl2
4 ml 5X Buffer with MgCl2
4 ml 5X Bufferw/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*
dNTPs
Pfu Polymerase
primers
DNA Template
MG Water
Cycling conditions
5X
5U/ul
10mM each
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**
dNTPs
Pfu Polymerase
primers
MgCl2
DNA Template
MG Water
5X
5U/ul
10mM each
1ug/ul each
50mM
-
-
1X
200uM
0.025U/ul
1.5-4.0 mM
-
-
50ng/ul each
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
45 sec
annealing (2)
extension
50-68°C
72°C
45 sec
45 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.
