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Sku: RT-PCR-146-2

Description

  • The 1-Step 2X Multiplex RT-PCR Master Mix is used for real-time qualitative and quantitative multiplex RT-PCR amplifications with TaqMan probes for up to four templates. 
  • The master mix is a premixed, 2X concentrated solution that has all the components except for gene-specific primers, probes and RNA templates.

Kit Characteristics

  • Direct-from-sample: enables expression analysis directly from cultured cells without RNA purification.

  • Fast preparation: 5-minute lysis including DNase treatment, versus 30–60 minutes for traditional RNA purification.

  • The kit is designed for multiplex RT-PCR with TaqMan probes.
  • For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which is a thermophilic type A polymerase, with optimal temperatures of 60-62°C.
  • The RTase is easily heat-inactivated at ≥90°C for 1min.
  • The RTase efficiently synthesizes complementary DNA strands on RNA templates from gene-specific primers, ≥1 unit per 20μL of reaction.
  • The RTase reversely-transcribes single digit copies of target RNA molecules consistently.
  • The kit also contains Taq-Probe DNA polymerase, specially engineered for TaqMan probes and generating S-shaped curves.
  • Up to four pairs of gene-specific primers can be applied in one reaction.
  • The concentrations of the primers and probes are variable depending on assay designs and thermocycling protocols (User Manual Table 1).
  • The preferable PCR product size is ≤150bp.
  • The kit has three formulations of ROX, Low ROX or High ROX concentration for your choice.

Kit Components

  • 2X Master Mix (2 × 1 mL / 5 × 1 mL / 10 mL for 200, 500, or 1000 × 20 μL reactions).

  • Lysis Buffer (2 × 10 mL / 1 × 50 mL / 1 × 100 mL).

  • User Manual.

Storage

Store at -20°C (stable up to 24 months with ≤10 freeze-thaw cycles). Transport at ≤4°C for up to 3 days.

Reaction Setup

20 μL Reaction: 10 μL Master Mix, primers, ≤4 μL RNA sample, water to 20 μL.
10 μL Reaction: 5 μL Master Mix, primers, ≤2 μL RNA sample, water to 10 μL.
Recommended amplicon size: ≤150 bp.

Compatible Instruments

Compatible with all major real-time PCR instruments, including but not limited to Applied Biosystems®, Bio-Rad®, Roche®, Qiagen®, Eppendorf®, and more.

Workflow


  1. 1.Harvest 10,000–100,000 cells.

  2. 2.Add lysis buffer and incubate for 5 minutes.

  3. 3.Heat at 95°C for 10 minutes.

  4. 4.Add lysate directly to RT-PCR setup.

  5. 5.Run RT-qPCR and analyze results.

Product Comparison

142: SYBR Green, No multiplexing, No probe system, Standard single-gene expression.

146: TaqMan, Yes multiplexing, Yes probe system, Multiplex TaqMan assays.

149: TaqMan, Yes (Super Multiplex), Yes probe system, Highly complex multi-target detection.

Product Manual

User Manual

When can I expect my order to ship?

Most orders are filled and shipped within 2-3 business days from the time they are received.

Our standard shipping usually take 2-5 days.

We also provide express shippping for time-sensitive deliveries. 

Email contact@biofargo.com if you have any requirements.

 

Terms and Conditions

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