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

Descripción general

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

Especificaciones y pureza

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

Estabilidad y almacenamiento

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

Condiciones de almacenamiento de almacenamiento

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

Enviado en

Ice chest + Ice pads

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Grado

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