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
2X Optimum RTL qPCR Master Mix is a NEW 2X premixed, ready-to-use solutions (GeneSpin proprietary formulation) containing new S∆Taq Pol, dNTPs, MgCl2 and stabilizers optimized for use in real time PCR amplification of DNA or cDNA.
2X Optimum RTL 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.
The 2X Optimum RTL qPCR Master Mix is supplemented with an inert blue dye and a separate Yellow Sample Buffer that contains a yellow dye. The qPCR reaction mix containing both components is green.
Endpoint PCR
cat. no.
QSTS-OLRSMMix200 RTL
QSTS-OLRSMMix500 RTL
QSTS-OLRSMMix1000 RTL
amount
note
5ml
2X conc. - low ROX
12.5ml
2X conc. - low ROX
25ml
2X conc. - low ROX
FOR RESEARCH USE ONLY
SHIPPING
Shipped in green ice.
STORAGE
Store at -20C°
avoid freeze/thaw cycles, store dark
Storage at 4 °C for up to 3 months possible.
SHELF LIFE
12 months
FORM
Liquid
CONCENTRATION
2X
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
2X
1X
10.0ul
25.0ul
primer for
10uM
100-400 nM
0.2 - 0.8 ul
0.5 - 2.0 ul
primer rev
DNA Template*
10uM
-
-
100-400 nM
0.2 - 0.8 ul
< 500ng
0.5 - 2.0 ul
<500ng
MG Water
-
-
up to 20ul
up to 50ul
*use 40X Yellow for diluition
REMEBER: The green color indicates that
the yellow sample A has been added into blue qPCR Master Mix
TEMPLATE and 40X Yellow assay set-up:
IN A FINAL qPCR REACTION OF 20ul
Template volume in 20ul
1ul
2ul
4ul
5ul
40X Yellow final conc. in template
20X
10X
5X
4X
DNA Template
50ul
25ul
12.5ul
10ul
i.e. for 5ul sample A in triplicate in 20ul reaction:
5ulx3 = 15ul sample A
15ul + 1.5ul of 40X Yellow = 16.5ul Yellow mix
Aliquot 5ul of mix for well.
REMEBER: The green color indicates that
the yellow sample A has been added into blue qPCR Master Mix
Cycling conditions:
Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.
denaturation
95°C
5min
denaturation (1)
annealing (2) and extension
95°C
55-68°C
10 sec
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.
SYBR® is a registered trademark of Invitrogen Corporation, Carlsbad, California, USA
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www.genespin.com
info@genespin.com
administration@pec.genespin.com
