2× Medino® Gold PCR Master Mix

Product2× Medino® Gold PCR Master Mix
Catalog No.LM201002-1
Storage conditionsThis product should be stored at -20℃ for 2 years.

Description

Product description
2×  Medino® Gold PCR Master Mix is a ready-to-use 2× premix solution, containing Medino® Gold High-Fidelity DNA Polymerase, dNTPs, and an optimized buffer system. The premix includes a heat-activated factor for Medino® Gold High-Fidelity DNA Polymerase, greatly enhancing the detection rate and specificity of the amplification. An extension factor added to the mix enables the enzyme to amplify long fragments, with target fragment lengths of up to 10 kb. This enzyme exhibits 5’→3’ DNA polymerase activity and 3’→5’ exonuclease activity, offering fidelity 52 times greater than that of Taq DNA Polymerase and six times that of regular Pfu DNA Polymerase. The amplification speed can reach 15 sec/kb. It is suitable for the amplification of complex templates and produces blunt-ended products. 2×  Medino® Gold PCR Master Mix has the advantages of being fast, simple, highly sensitive, highly specific, and stable. The reaction system only needs the addition of primers and template, and can be amplified using a two-step procedure, simplifying experimental steps and saving time. This product does not contain dye; thus, PCR products must be added to Loading buffer before electrophoresis. Additionally, this product contains a special protective agent, allowing the premix to maintain stable activity after repeated freeze-thaw cycles.
Components
Cat.No. LM201002
Size1 mL/5×1 mL
Instructions
  1. Recommended PCR reaction systems.
ComponentsVolume (μL)Final concentration
ddH2OUp to 50
Templatex
Forward Primer 10 μmol/L2.50.5 μmol/L
Reverse Primer 10 μmol/L2.50.5 μmol/L
2 ×Medino® Gold PCR Master Mix25
[Note]:
  1. a) Polymerase final concentration: 1× containing 1 U / 50 μL polymerase
  2. b) Primer: The final concentration of primer in the PCR reaction system is 0.2 – 1 μM, recommended as 0.5 μM.
  3. c) Mg2 +, dNTP, final concentration: 1.5 mM Mg2 + and 200 μM.
  4. d) Recommended usage for different templates (50 μL reaction system):
TemplateThe amplified fragment was 1 kb – 10 kb
Genome DNA50 ng-200 ng
Plasmid or viral DNA10 pg-20 ng
cDNA1-5 μL (not more than 1 / 10 of total volume of PCR reaction)
  1. Reaction program
Cycle stepTemp.TimeCycles
Initial denaturation98℃3min1
Denaturation98℃10 sec30-35
Annealing60℃20 sec
Extension72℃30 sec/kb
Final extension72℃5 min1
[Note]:
  1. Pre-denaturation temperature and time: recommended temperature: 98℃, time: 3 min, high GC content template 5-10 min.
  2. Annealing temperature and time: recommended temperature: 60℃, or according to the need, set up a temperature gradient to explore the optimal temperature of primer annealing. The recommended annealing time is set to 20 sec, which can be adjusted within 10-30 sec. Too long an annealing time may cause the dispersion of the amplification product on the glu
  3. Extended temperature and time: Recommended temperature: 72℃. Time: 30 sec/kb, and the complex template can be extended to 60 sec/kb according to the actual situation.
  4. d) Amplification product: Please place the PCR amplified product in -20℃ to prevent the enzyme from degrading the amplified product. The amplified products are shown at the flat ends.
Notes
  1. For your safety and health, please wear a lab coat and disposable gloves during
  2. This product is for research use only!
  3. After thawing, Master Mix may precipitate. Before use, vortex and mix until the precipitate disappears, without affecting the performance of the reagent.
Documents

Reviews

There are no reviews yet.

Be the first to review “2× Medino® Gold PCR Master Mix”

Your email address will not be published. Required fields are marked *