Sanshibio 2× Taq PCR Master Mix(con colorante)1 ml

$3.50

Spedizione USD 45 - Gratuito per USD 300

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Descrizione

2× Taq PCR Master Mix(with dye)

oggetto numeroE002 Specificazione:1mL Storage: Store at -20°C

Introduzione al prodotto

The 2x Taq PCR Master Mix (con colorante) is a premixed solution at twice the concentration of DNA Polymerase, PCR Buffer, dNTP Mix, and indicators. When using this product, simply add the template and primers to the premixed solution, and complete the volume with ddH2O for the PCR reaction. This simplifies the operational steps and reduces the risk of contamination during PCR procedures. Inoltre, the premixed solution contains the necessary dye reagents for gel electrophoresis, enabling direct gel electrophoresis after PCR. This product offers high sensitivity, strong specificity, and a wide linear range, allowing for more accurate quantification of target genes. The PCR products amplified using this product have an added A base at the 3end, facilitating cloning into T vectors.

Contenuto del prodotto:

Componenti E002
2× Taq PCR Master Mix(with dye) 1ml

Magazzinaggio:

Store at -20°C, with a minimum shelf life of 12 mesi.

Controllo di qualità:

This product has undergone quality testing and is free from endonuclease activity, exonuclease activity, and ribonuclease contamination. Residual host genomic DNA is below 10 copies.

Product Usage:

Amplification of DNA using PCR method; DNA sequence determination.

Usage Instructions:

  • Allow all required PCR reaction solutions to equilibrate to room temperature until fully dissolved. Thoroughly mix the solutions and set up the PCR reaction system on ice.
  • It is recommended to set up the PCR reaction system on an ice bath or in a cooler, following the recommended reaction system for reference.
  • Thoroughly pipette and mix the prepared reaction system using a pipette, seal the PCR tubes with caps, label them appropriately, and briefly centrifuge at room temperature.
  • Place the prepared PCR tubes into the Macchina per PCR, set the reaction conditions, and initiate the PCR reaction.

Recommended Reaction System:

Reagenti 25µL System Volume Final Concentration
2× Taq PCR Master Mix(with dye) 12.5μL
Primer I (10µM) 0.5-2.5μL 0.2-1.0μm
Primer II (10µM) 0.5-2.5μL 0.2-1.0μm
Template DNA 1μL as required
ddH2O Up to 25μL

Reaction Conditions:

Under normal circumstances, a two-step method can be used for the reaction. If the two-step amplification is not satisfactory, a three-step method can be employed to set up the PCR reaction program. Considering the uncertainty of target gene abundance in cDNA templates, it is recommended to use 35-38 cycles for the initial amplification and adjust the number of cycles based on the amplification results.

Method/Steps Time setting Cycles
95℃ (Pre-denaturation) 2-5min 1
95℃ (Denaturazione) 10sec 28-40Cycles

(Plasmid or Genomic Template:28-35cicli;

cDNA Template:30-40cycles)

55℃-60℃ (Ricottura) 30sec
72℃ (Estensione) 1min / 1-3kb
72℃ (Final Extension) 5-10min 1
4℃/16℃(Hold)

Nota: Reaction conditions can be adjusted and optimized according to actual requirements.

Precauzioni:

  1. Extension time can be adjusted based on factors such as the length and GC content of the PCR product. The extension time per kb of the product is closely related to the complexity of the template: simple templates require 20 secondi, typical templates require 30 secondi, and complex templates require 1 minuto.
  2. The setup of the PCR reaction should be tailored to different conditions such as template, primer, length of PCR product, and GC content. The final concentration of primers typically falls within the range of 0.2-1.0μM. DNA template concentration can be adjusted accordingly. For complex templates or high GC content, it’s recommended to prolong pre-denaturation/denaturation or extension times and increase denaturation/annealing temperatures.
  3. Use designated areas and pipettes before and after amplification, wear gloves, and change them frequently. After completing the PCR reaction, do not open the reaction tubes immediately. Allow them to cool sufficiently at 4°C or -20°C before opening to minimize the risk of PCR product contamination to the laboratory environment.

*For OEM, please contact to our salesperson

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