Ultra - High Fidelity NGS Multiplex PCR Master Mix II , 2X

Ultra - High Fidelity NGS Multiplex PCR Master Mix II , 2X

Product Details:

Place of Origin: China
Brand Name: GDSBio
Certification: ISO9001, ISO13485
Model Number: NM2001/NM2002/NM2003

Payment & Shipping Terms:

Minimum Order Quantity: 1 bag
Packaging Details: small package or bulk distribute or OEM
Delivery Time: 8 work days
Payment Terms: L/C, D/A, D/P, T/T, Western Union, MoneyGram
Supply Ability: 100 Bag/Bags per Day
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Detail Information

Stock: Yes Cat. No.: NM2001/NM2002/NM2003
Specification: 40 Reactions/400 Reactions/2000 Reactions Appearance: Clear Liquid
Logo Printing: With Logo Printing Transport Package: Packing
Storage Conditions: -20°C
High Light:

NGS Multiplex PCR Master Mix II

,

PCR Master Mix II

Product Description

NGS Multiplex PCR Master Mix II, 2X

 

The NGS Multiplex PCR Master Mix II contains all of the components for NGS multiplex PCR (except for primers and templates) in a single tube, including chemically modified ultra-high fidelity DNA Polymerase, which has 100X amplification fidelity of Taq DNA Polymerase. The PCR products have an ultra-low error rates, and is very suitable for the detection of tumor ctDNA and MRD.

 

Cat. No. Contents Storage Conditions
NM2001 NGS Multiplex PCR Master Mix II, 2X, 40 reactions

 

Store unopened at –15°C to –25°C until the expiration date on the label.

After opening, the master mix may be stored at –15°C to –25°C until the expiration date on the label, or at 4°C for up to 30 days.

The GC Enhancer must be kept at –15°C to –25°C.

  • NGS Multiplex PCR Master Mix II, 2X (1 × 1 mL)
  • GC Enhancer (1 × 0.25 mL)
NM2002 NGS Multiplex PCR Master Mix II, 2X, 400 reactions
  • NGS Multiplex PCR Master Mix II, 2X (10 × 1 mL)
  • GC Enhancer (2 × 1 mL)
NM2003 NGS Multiplex PCR Master Mix II, 2X, 2000 reactions
  • NGS Multiplex PCR Master Mix II, 2X (5 × 10 mL)
  • GC Enhancer (1 × 10 mL)

 

 

Protocol

Note: Before setting up the PCR reactions, prepare a Primer Mix II with 0.5 µM of each primer.

 

Prepare the PCR Reaction Mix

1. Allow all reagents to thaw completely. Mix gently by inverting the tube. Spin briefly. Put all reagents on ice.

2. Using a 96-well optical reaction plate, add the following to one well per sample:

 

Component Volume or Mass Final Concentration
NGS Multiplex PCR Master Mix II, 2X 25 µL 1X
Primer Mix II(0.5 µM each) 5 µL 50 nM each primer[1]
Template DNA 0.1–0.2 µg 2–4 ng/µL
GC Enhancer 0 or 6 µL [2] 0 or 12%
Nuclease-free water Adjust to 50 µL n/a

[1] The final concentration of each primer in a typical PCR is between 0.05-0.4 μM. In most cases, a final concentration of 0.15 μM gives satisfactory results. Increasing the primer concentration up to 0.4 μM may increase the yield.

[2] Use GC Enhancer only when high GC content targets cannot be amplified under standard conditions.

 

3. Seal the reaction plate with Clear Adhesive Film.

 

Amplify DNA for Analysis

Choose an amplification protocol based on your analysis method

 

Amplify for analysis by agarose gel electrophoresis

 

1. Configure the run method as outlined in your instrument’s user manual. Use the following parameters:

Three-step reaction system:

 

Step Time Temperature (°C)
Hold 1 min 98
25-35 Cycles 15 sec 98
30 sec 58
30 sec 72
Hold 2 min 72
Hold ¥ 4

Two-step reaction system(Tm>60℃):

Step Time Temperature (°C)
Hold 1 min 98
25-35 Cycles 15 sec 98
60 sec 60
Hold 2 min 72
Hold ¥ 4

2. Mix well and briefly spin the reaction plate.

3. Load the reaction plate into the instrument, and start the run. See your instrument’s user manual for detailed instructions on how to load and run the plate.

4. Analyze the data according to the instructions in the user manual for your gel electrophoresis instrument.

 

Amplify for CfDNA Sequencing

 

Component Volume
NGS Multiplex PCR Master Mix II, 2X 12.5 µL
Primer Mix II 2 µL
cfDNA X µL
Nuclease-free water To 25 µL

 

1. Configure the run method as outlined in your instrument’s user manual. Use the following parameters:

Three-step reaction system:

 

Step Time Temperature (°C)
Hold 1 min 98
25-35 Cycles 15 sec 98
30 sec 58
30 sec 72
Hold 2 min 72
Hold ¥ 4

Two-step reaction system(Tm>60℃):

Step Time Temperature (°C)
Hold 1 min 98
25-35 Cycles 15 sec 98
60 sec 60
Hold 2 min 72
Hold ¥ 4

2. Mix well and briefly spin the reaction plate.

3. Load the reaction plate into the instrument, and start the run. See your instrument’s user manual for detailed instructions on how to load and run the plate.

Ultra - High Fidelity NGS Multiplex PCR Master Mix II , 2X 0

GDSBio is a high-tech company focused on the development, production and marketing of quality life science products. The company has a complete product line, with PCR technology as the core, focusing on ordinary PCR, fluorescent quantitative PCR, NGS library construction, nucleic acid electrophoresis and other molecular biology technologies, and has developed molecular scientific research reagents, molecular in vitro diagnostic raw materials, nucleic acid extraction and detection reagents and other products.

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