MB qPCR OptSYBR RTL

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 fluorescein for dynamic well factor collection. The addiction of fluorescein 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.

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Optimum qPCR Master Mix

cat. no.

amount

note

QSTS-OSMMix200 RTL

QSTS-OSMMix500 RTL

5ml

2X blue + 4.0ml 40X Yellow

2X blue + 1.5ml 40X Yellow

QSTS-OSMMix1000 RTL

25ml

2X blue + 8.0ml 40X Yellow

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 blue

CONCENTRATION

12.5ml

MG Water

> 2X Optimum RTL qPCRMaster Mix with SYBR® Green I

qPCR

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

2X Master Mix

primer for

primer rev

DNA Template

stock conc.

10uM

final conc.

100-400 nM

20ul reaction

10.0ul

0.2 - 0.8 ul

< 500ng

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

denaturation

denaturation (1)

annealing (2) and extension

95°C

55-68°C

5min

10 sec

30 sec

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.

50ul reaction

25.0ul

0.5 - 2.0 ul

<500ng

MG Water

up to 20ul

up to 50ul

melt curve

from 65°C to 95°C

0.5°C/5sec

SYBR® is a registered trademark of Invitrogen Corporation, Carlsbad, California, USA

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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

molecular

biology

yellow template*

*use 40X Yellow for diluition

Template volume in 20ul

40X Yellow final conc. in template

DNA Template

1ul

20X

50ul

2ul

10X

25ul

4ul

12.5ul

5ul

10ul

Volume 40X Yellow in 100ul template

TEMPLATE and 40X Yellow assay set-up:

IN A FINAL qPCR REACTION OF 20ul

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.

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