Medino® V Universal Multiplex One Step RT-qPCR Probe Kit

 

Cat.No. LM602006
Size100 T / 500 T / 1000 T / 10000 T
Storage and shipping1. The product is shipped with ice pack.

2.  The product can be stored at -15℃ ~ -25℃ for 18 months.

Application equipment

 

ABI 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™, StepOne Plus™,ABI 7500, 7500 Fast, ViiA™7, QuantStudio ™ 3 and 5, QuantStudio™ 6,7,12k Flex

Stratagene MX3000P™, MX3005P™, MX4000P™

Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, MyiQ™, MiniOpticon™, Opticon®, Opticon® 2, 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

Thermo Scientific PikoReal Cycler; Cepheid SmartCycler®; Illumina Eco qPCR

Product description

Medino® V Universal Multiplex One Step RT-qPCR Probe Kit is a kit for performing multiplex quantitative PCR reactions using RNA as a template. During the experiment, reverse transcription and quantitative PCR were performed in the same reaction tube, which simplified the experimental operation and reduced the risk of contamination.

This kit uses heat-stable Medino® V Reverse Transcriptase to efficiently synthesize first-strand cDNA and Medino® HotStart Taq DNA Polymerase for quantitative amplification. This kit mainly contains optimized MP Buffer, Enzymes Mix, etc. The buffer already contains Mg 2+ and dNTPs, etc., and has added factors that effectively inhibit non-specific PCR amplification and factors that improve the amplification efficiency of multiplex qPCR reactions, which can Perform up to four-plex reactions while ensuring primer amplification efficiency.

Components

CompontNo.NameFormat
100T500T1000T10000T
A2×MP Buffer1.25  mL6.25 mL12.5 mL125 mL
BEnzyme Mix100μL500 μL1 mL10 mL

[Note]:

  1. 2×MP Buffer are the abbreviation for Multiplex One Step RT-qPCR Probe Buffer.
  2. Enzyme Mix mainly contains heat-resistant V Reverse Transcriptase and HotStart Taq DNA Polymerase.

 

Operate

Reaction System

Components Volume μLFinal concentration
2×MP Buffer12.51 ×
Enzyme Mix1 
Primer mix (10 μM)1 each0.4 μM
Probe mix (10 μM)0.5 each0.2 μM
Template1 – 10
RNase free H20Up to 25

[Note]:  Be sure to mix thoroughly before use, avoiding excessive bubbles caused by vigorous shaking.

  1. Primer concentration: Primer Mix contains multiple pairs of primers and can be adjusted between 0.1-1.0 μM according to the situation.
  2. Probe concentration: Probe Mix contains multiple probes with different fluorescence signals, and the concentration of each probe can be adjusted between 50-300 nM according to specific conditions.
  3. Template dilution: The sensitivity of qPCR is extremely high. It is recommended to dilute the template and control the Ct value between 20-35.
  4. Reaction system: 25-50 μL is recommended to ensure the effectiveness and reproducibility of target gene amplification.
  5. System preparation: Please prepare in a clean workbench, and use pipette tips and reaction tubes without nuclease residues; it is recommended to use pipette tips with filter elements; avoid cross-contamination and aerosol contamination.

Reference reaction program

Cycle stepTemp.TimeCycles
Reverse transcription50 °C a)10 min1
Initial denaturation95 °C5 min1
Amplification reaction95 °C15 sec45
60 °Cb)30 sec c)

[Note]:

  1. Reverse transcription: 42℃ or 50℃ can be used.
  2. Amplification reaction: The amplification reaction temperature is adjusted according to the designed primer Tm value.
  3. Fluorescence signal collection: Different qPCR detection instruments require different fluorescence signal collection times. Please set according to the minimum time limit.

Notes

  1. For your safety and health, please wear lab coats and disposable gloves for operation.
  2. Please use RNase free consumables.
  3. This product is for research use ONLY
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