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Super Green qPCR mix (~Sybr Green qPCR mix，None/Low/High ROX & Universal)

Super Green qPCR Master Mix is a ready-to-use cocktail containing all components except primers and template.
The 2X master mix contains Taq DNA polymerase, dNTPs, MgCl2, SYBR Green I, Rox or No Rox and stablizers. In the formulation, for HotStart, Taq DNA
Polymerase is chemically modified and its activity is completely blocked until the first denaturation step in PCR program.
This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency. For easy and avoiding potential error manipulation, the products are provided in three formats: no Rox, low Rox, high Rox.

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市场价

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浏览量:

1000

产品编号

021-50T/150元; 200T/500元

所属分类

qPCR reagents

数量

库存:



产品描述

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Super Green qPCR mix (~Sybr Green qPCR mix)

(None/Low/High ROX & Universal)

Super Green qPCR Master Mix is a ready-to-use cocktail containing all components except primers and template. The 2X master mix contains Taq DNA polymerase, dNTPs, MgCl₂, SYBR Green I, Rox or No Rox and stablizers. In the formulation, for HotStart, Taq DNA Polymerase is chemically modified and its activity is completely blocked until the first denaturation step in PCR program. This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency. For easy and avoiding potential error manipulation, the products are provided in three formats: no Rox, low Rox, high Rox. Customers could choose suitable product format based on their qPCR instruments used.

Product Components:

Cat. Num.	Size	Cat. Num.	Size
F021-NOROX-1	1 ml	F021-NOROX -5	4X1.25 ml
F021-LOWROX-1	1 ml	F021-LOWROX -5	4X1.25 ml
F021-HIGHROX-1	1 ml	F021-LOWROX 5	4X1.25 ml

F021-NOROX contains no ROX, F021-LOWROX contains low ROX，F021-HIGHROX contains high ROX.

Following table is helpful for choosing right product formats

No ROX	Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4; Cepheid SmartCycler®; Eppendorf Mastercycler® ep realplex, realplex 2 s; Illumina Eco qPCR; Qiagen/Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene 6000;Roche Applied Science LightCyclerTM 480; Thermo Scientific PikoReal Cycler
Low ROX	Applied Biosystems 7500, 7500 Fast, ViiA7; Stratagene MX4000, MX3005P, MX3000P
High ROX	Applied Biosystems 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast; StepOne, StepOnePlus.

Storage Condition:

Store at -20℃avoiding light, the master mix should be not stored at 4℃ for longer time after being thawed. The mix can tolerate 30 cycles freeze and thaw without affecting Ct value for sample analysis, keep at -20℃ if no plan to use in certain time. Please blending the master mix before aliquot.

Experimental Procedure

1. Set up reaction in qPCR tube as follow:

2 × Super SYBR qPCR Master Mix	10.0 µl
Primer1 (10 µM)	0.4 µl
Primer2 (10 µM)	0.4 µl
Template DNA/cDNA	X µl
ddH2O

Quantity of components in the reaction could be adjusted as needed to achieve better results:

1) Generally 0.2 μM primer concentration is suitable, but when results are not satisfied, trying primer concentration between 0.1-1.0 μM range

2) qPCR method is very sensitive, accuracy of added temple is essential, recommending using diluted templates to reduce to increase aliquot accuracy, and the result is reproducible.

3) If templates are undiluted cDNA from standard reverse transcription reaction, the volume of template is no more than 10% of the reaction volume.

2. Running qPCR reaction as following

Stage 1

Pre-denatuea

Reps: 1

95 ℃

5-10 min

Stage 2

Cyclingb

Reps: 40

95 ℃

60 ℃

10 sec

30 sec

Stage 3

Melting curvec

Reps: 1

95 ℃

60 ℃

95 ℃

15 sec

60 sec

15 sec

2.1Pre-denature condition is suitable for most of reactions, if templates are complicated, extend to 10 min.

2.2for less 300 bp fragment amplification, 30 second extending time is enough, for large than 300 bp fragment amplification, 60 second extending time is recommended.

2.3Melting curve collecting program depends on instrument’s model, please choose acquiescence for the model

Quality Control

Purity detection：all components are analyzed without exo- and endo-nulease and nuclear acid.

Optimizing reaction

Best reaction condition should have following characteristic: single melting curve, amplification efficiency is almost 100%, lower Ct value (high amplification efficiency), if reaction is not as expected under acquiescence condition, reaction condition could be optimized as following ways.

1. Primer concentration and reaction: when primer concentration is between 0.1～1.0 μM, higher primer concentration leads non-specific amplification, but amplification efficiency is increased.

2. Amplification program and reaction: To increase amplification specificity, increase annealing temperature and extending amplification time

3. To increase amplification efficacy, change two step amplification to three step and increase extending time.

NOTE: Always wear lab coats, gloves and goggles when working with our products although they are low-risk chemicals for R&D only.

特别提醒：

1) 本公司所有产品仅限于专业人员用于生命科学研究，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅。

2) 本公司所有产品必须由合格专业技术人员操作同时佩戴口罩/手套/实验服并遵守生物实验室安全操作规程！

Description

Name	Super Green qPCR mix (~Sybr Green qPCR mix，None/Low/High ROX & Universal)
CAT#	021-50T; 021-200T	CAS#	N/A
Storage#	-20°C Sealed & desiccated & Minimized light exposure & Minimized Freezing And Thawing	Shelf Life#	24 months
Ex(nm)#	497	Em(nm)#	525
MW#	N/A	Solvent#	N/A

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