2X Optimum qPCR Master Mix is a NEW 2X premixed, ready-to-use solutions (GeneSpin proprietary formulation) containing new OPTIMUM Taq Pol, dNTPs, MgCl2 and stabilizers optimized for use in real time PCR amplification of DNA or cDNA.
2X Optimum Master Mixes offers more stability, reproducibility and tolerance to PCR inhibitors, and contain ROX for dynamic well factor collection. The addiction of ROX has no effect on the PCR reaction efficiency or sensitivity of detection.
cat. no.
cat. no.
amount
note
QSTS-OLRSMMix 200
QSTS-OLRSMMix 500
5 ml
12.5 ml
2X conc. - low ROX
2X conc. - low ROX
QSTS-OLRSMMix 1000
5x5 ml
2X conc. - low ROX
FOR RESEARCH USE ONLY
SHIPPING
Shipped in green ice.
FORM
Liquid
SHELF LIFE
12 months
STORAGE
Store at -20°C
CONCENTRATION
2X conc.
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.
20ul reaction
50ul reaction
2X Master Mix
primer for
primer rev
DNA Template
2X
-
10 uM
10 uM
1X
100-400 nM
100-400 nM
-
10.0 ul
0.2-0.8 ul
0.2-0.8 ul
< 500ng
25.0 ul
0.5-2.0 ul
0.5-2.0 ul
< 500ng
MG Water
-
-
up to 20ul
up to 50ul
Cycling conditions
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)
and extension
55-68°C
30 sec
melt curve
from 65°C
to 95°C
0.5°C/5sec
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.
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