UltraBio™ Multiplex Amplification Mix - BioReagent,DNase, RNase free,PCR Reagent,Suitable for molecular biology,for DNA and RNA applications,2.5μL/T

Cat. No.: M751562

GRADE & PURITY Suitable for molecular biology, BioReagent, DNase, RNase free, PCR Reagent, for DNA and RNA applications, 2.5μL/T

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Application

Multiplex PCR, PCR

Overview

UltraBio™ Multiple Amplification Mix is a universal multiplex amplification reagent that can be used for constructing multiplex amplicon libraries for microbial/genetic mutation detection. It is compatible with ultra-multiplex amplification, with a primer number range of 1 to 1500, amplicon length range of 100 to 1800 bp, GC content range of 13% to 75%, and a minimum template input amount of 1 ng. 

   The reagent undergoes strict background microbe quality control to minimize false positives from non-specific amplification. 

   Rigorous functional validation ensures high stability and reproducibility, making it ideal for NGS-based multiplex targeted amplification in microbial/genetic mutation detection.

Catalog	Component	24T	96T	Storage
M751562	UltraBio™ Multiplex Amplification Mix	60 μL	240 μL	-20℃

Self-Supplied Materials

1. Instruments

    PCR thermal cycler, Pipettes, Magnetic stand, Microcentrifuge, Vortex mixer, Agarose gel electrophoresis system, UV gel imaging system, Agilent Bioanalyzer 2100 (or equivalent).

2. Reagents

    Agarose, 1× TAE buffer, Nucleic acid dye, DNA marker, Ampure XP beads (or equivalent), Freshly prepared 80% ethanol, Nuclease-free water.

3. Consumables

    1.5 mL microcentrifuge tubes, 0.2 mL PCR tubes, DNase/RNase-free pipette tips.

Protocol

I. Multiplex Amplification & Product Verification

1. Reagent Preparation

   • Thaw UltraBio™ Multiplex Amplification Mix on ice.

   • Vortex briefly and centrifuge before use.

2. Reaction Setup (10 μL Standard Volume)

3. Component	Volume per Reaction	Final Concentration
   UltraBio™ Multiplex Amplification mix	2.5 μL	4x
   Primer Mix	1 μL	20nM (2.5~100nM)
   Template DNA	X μL	10ng (1~100ng)
   Nuclease-free water	Up to10 μL	/
   Total	10 μL	/

Note:

• Total reaction volume can be scaled to 10–50 μL (adjust mix proportionally).

• Primer concentration and annealing temperature (T) should be optimized for each panel.

1. Mixing & Centrifugation

   • Vortex gently and centrifuge briefly to collect liquid.

2. PCR Cycling Program

*T: Annealing temperature depends on primer panel (optimize empirically).
Cycle number: Adjust based on template input (default: 25 cycles).

1. Product QC: Analyze amplification success via 2% agarose gel electrophoresis or Agilent 2100 Bioanalyzer.

II. Amplicon Purification (Ampure XP Beads)

1. Bead Equilibration

   • Warm Ampure XP beads to room temperature; vortex thoroughly.

2. Bead Binding

   • Dilute PCR products to 50 μL with nuclease-free water.

   • Add 60 μL beads (1.2× ratio), vortex, and incubate 5 min at RT.

3. Wash Steps

   • Place tubes on a magnetic stand for 3–5 min; discard supernatant.

   • Wash twice with 200 μL fresh 80% ethanol (30 sec per wash).

   • Air-dry beads for 2–3 min until matte.

4. Elution

   • Resuspend beads in 22 μL nuclease-free water; incubate 2 min at RT.

   • Transfer 20 μL supernatant to a new PCR tube.

   • Store purified products at –20°C.
     

Specifications

Specifications & Purity

BioReagent, DNase, RNase free, PCR Reagent, Suitable for molecular biology, for DNA and RNA applications, 2.5μL/T

Stability And Storage

Store at -20℃ long term (12 months). Avoid freeze/thaw cycle. Upon receipt, it is recommended to aliquot.

Storage

Store at -20°C, Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent, DNase, RNase free, for DNA and RNA applications, PCR Reagent, Suitable for molecular biology