Medino ® Universal Multiplex qPCR Master Mix (Probe Based)

 

Catalog No.LM602002
Storage conditionsThe product should be stored at -25°C  – 15°C  for  2 year.
Hot StartBuilt-in hot start
Detection methodPrimer-probe detection
PCR methodqPCR
PolymeraseTaq DNA polymerase
Type of sampleDNA
Application equipmentApplied  Biosystems:   5700,   7000,   7300,   7700, 7900HT Fast,  StepOne™,  Stepone  Plus™,  7500, 7500   Fast,   ViiA™7,   Quantstudio™   3   and   5, Quantstudio™ 6,7,12k Flex;

Bio-Rad: CFX96, CFX384, iCycleriQ, iQ5, MyiQ, Miniopticon, Opticon, Opticon2, Chromo4;

Eppendorf: Mastercycler ep realplex, realplex 2 s;

Qiagen: Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene    6000;    Roche    Applied     Science: LightCycler 480, LightCycler 2.0; Lightcycler 96;

Stratagene:          MX3000P™,          MX3005P™, MX4000P™;

Thermo   Scientific:   PikoReal   Cycler;   Cepheid: SmartCycler; Illumina: Eco qPCR.

Product description

Universal Multiplex qPCR Master Mix (Probe Based) is a pre-solution for 2 × real- time quantitative PCR amplification characterized by high sensitivity and specificity, is blue in color, and has the effect of sample addition. This product is a 2 X Mix pre-mixed reagent that enables up to  four fluorescent quantitative PCR reactions in  a  single reaction well. This product contains the genetically modified antibody method to hot- start Taq enzyme, greatly improving the amplification sensitivity and specificity. At the same time, this product has deeply optimized the multi-reaction buffer, which can improve  the  amplification   efficiency  of  the  reaction   and  promote   the  effective amplification of low-concentration templates. This product can be used for genotyping and multiplex quantitative analysis.

Components

Components

No.

Name11211ES0311211ES0811211ES09
LM602002

UniversaI Multiplex

qPCR Master

Mix (Probe Based)

1  mL5 x 1 mL5 mL

Components

No.

Name11211ES2011211ES6011211ES61
LM602002

UniversaI Multiplex

qPCR MasterMix

(Probe Based)

20 mL100 x 1 mL100 mL

Instructions

1.Reaction System

ComponentsVolume (μL)Final Concentration
2 × Universal TaqMan multiplex qPCR master mix12.51 x
Primer mix (10 μmol/L)x0.1-0.5 μmol/L
Probe mix (10 μmol/L)x50-250 nmol/L
Rox reference dye0.41 x
Template DNA/cDNA1- 10
ddH2Oup to 25

[Note]: Mix thoroughly before use to avoid excessive bubbles from vigorous shaking.

  1. a) Primer concentration: Primer Mix contains multiple pairs of primers, usually each primer at a final concentration of 2 μmol/L and can also be adjusted between 0.1 and 0.5 μmol/L as appropriate.
  2. b) Probe concentration: Probe   Mix   contains   multiple   probes   with   different fluorescence signals, and the concentration of each probe can be adjusted between 50 and 250 nmol/L according to specific situation.
  3. c) Rox dye reference: It is used on Real Time PCR amplification instrument such as Applied Biosystems to correct the error of fluorescence signal generated between wells; this product does not contain Rox dye
  4. d) Template dilution: qPCR is highly sensitive, and it is recommended to dilute the template for use. If the template is a cDNA stock solution, the template volume should not exceed 1/10 of the total volume.
  5. e) Reaction system: 25 μL, 30 μL or 50 μL is recommended to ensure the effectiveness and repeatability of target gene amplifica
  6. f) System preparation: Please prepare in the super clean bench, and use the tips and reaction tubes without nuclease residue; it is recommended to use the tips with filter Avoid cross contamination and aerosol contamination.

Reaction program

Cyclestep Temp.   Time      Cycles
Initial denaturation95°C5 min1
Denaturation95°C15 s45
Annealing/Extension60°C30 s

[Note]:

  1. a) Annealing/Extension: The  temperature  and  time  can  be   appropriately adjusted according to the designed primer Tm value.
  2. b) Fluorescence signal acquisition: The fluorescence signal acquisition time required for different qPCR  detection  instruments  is  different,  please  set  according  to  the minimum time limit. The time of several common instruments is set as follows:

20 sec: Applied Biosystems TT00, T900HT, T500 Fast

31 sec: Applied Biosystems 7300

32 sec: Applied Biosystems 7500

 

Notes

Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety!

This product is for scientific research purposes only.

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