2× Medino® PCR Master Mix(for multiplex PCR)

Product2× Medino® PCR Master Mix(for multiplex PCR)
Catalog No.LM201012-1
Storage conditionsThis product should be stored at -20℃ for 2 years.

Description

Product description

2× Medino® PCR Master Mix (for Multiplex PCR) is developed based on a new generation of high-fidelity DNA, polymerase Canace, and is specifically optimized for multiplex PCR library construction applications in the field of high-throughput sequencing. The Canace enzyme is based on Pyrococcusfuriosis DNA Polymerase and is modified by genetic engineering. The protein structure contains the 5 ′-3 ′ polymerase domain and the modified 3 ′- 5 ′ exonuclease domain. This combination greatly improves the heat resistance and fidelity of the enzyme. It is 98℃, 52 times more fidelity than Taq DNA polymerase and 6 times more fidelity than common Pfu DNA polymerase. Unique elongation factors were added to the enzyme solution and the amplification was up to 15 sec/kb. This product is a 2 mix PCR mixture, with a large volume range of added templates and primers, making your experiment more flexible (another 5 mix is available).

Components

Cat.No.LM201012-1
Size1 mL/500mL

Instructions

  1. Recommended PCR reaction systems.
ComponentsVolume (μL)Final concentration
2× Medino® PCR Master Mix (for multiplex PCR)25
Templatex
Primer mix (X μM)x
ddH2OUp to 50
[Note]:
  1. a) Please mix each components before use.
  2. b) Primers and template: The first round of multiplex PCR for target enrichment, in which the primers    and template conditions need to be explored. The concentration of the second round of multiplex           PCR is recommended: 10 μM primer plus about 2 μL.
  1. Reaction program
Cycle stepTemp.TimeCycles
Initial denaturation98℃30 sec1
Denaturation98℃15 sec25-35
Annealing60℃30 sec
Extension72℃1 min
Final extension72℃5 min1
[Note]:
  1. a) Annealing temperature and time: 60℃ is recommended, and a temperature gradient can also be set    up to explore the optimal temperature of primer annealing as needed. The recommended annealing            time is set to 30 sec, which can be adjusted within 10-30 sec. Too long an annealing time may cause       the dispersion of the amplification product on the glue.b)   The extension time of 10 sec/kb can             amplify most of the target fragments below 8 kb, and most of                them below 2 kb at 3-5 sec/kb. If the               amplification efficiency is low, the time can be appropriately              extended to 20-30 sec/kb, and should              not exceed 60 sec/kb.

Notes

  1. a) For your safety and health, please wear a lab coat and disposable gloves during operation.
  2. b) This product is for research use only!
  3. c) It is worth noting that the multiple PCR library building experiment itself is difficult, which usually needs to be optimized. If you encounter difficulties when using this product, you can contact our               technical support. We have multiple optimization schemes, which can help you to better build the     experimental system.

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