Multiplex PCR MasterMix (UNG)
2024-10-25
T4 Polynucleotide Kinase
2024-10-25Product Number: PCK239
Shipping and Storage
Store at -20℃ and transport on dry ice.
Components
| Component | PCK239 24rxns | PCK239 96rxns |
| 10×End Repair Buffer | 200μl | 800μl |
| End Repair Enzyme Mix | 48μl | 192μl |
| Ligation and Nick Repair Buffer | 400μl | 2×800μl |
| T4 DNA Ligase | 48μl | 192μl |
| Bst DNA Polymerase | 48μl | 192μl |
| 2×HiFidelity PCR Mix | 600μl | 2×1.2ml |
| 10×Primer Mix (5μM each) | 150μl | 600μl |
Description
The NGS Fast DNA Library Prep kit for Ion Torrent provides the enzyme premix system and reaction buffer required for constructing DNA libraries, including all components except connectors. The prepared library can be used for Ion Torrent PGM Sequencing of IonPoton second-generation sequencing platform. Compared with conventional library building methods, this kit combines multiple steps and omits multiple purification steps, significantly reducing the minimum requirement for starting template DNA and shortening the library construction time. In addition, the reagent kit uses high fidelity DNA polymerase for library enrichment and preference free PCR amplification, expanding the coverage area of the sequence and efficiently preparing DNA libraries for the Ion torrent second-generation sequencing platform.
Self provided instruments, reagents, and consumables
- Magnetic frame
- DNA purification and recovery kit: It is recommended to use the Magnetic Bear DNA Purification Kit (for NGS Size Selection) (DNK2508).
- Sample connector primer kit.
- Anhydrous ethanol, EB (10 mMTris HCl, pH 8.0),Deionized water (pH between 7.0 and 8.0).
- Reaction tube: It is recommended to use a low adsorption PCR tube and a 1.5 ml centrifuge tube;
- Gun tip: It is recommended to use high-quality filtering gun tips to prevent contamination of reagent kits and library samples.
Preparation and important precautions before the experiment
- To avoid repeated freezing and thawing of the buffer in the reagent kit, it is recommended to package and store the buffer for the first time of use. Enzymes should be stored at -20℃ as soon as possible after use,
- PCR products are prone to contamination due to improper operation, resulting in inaccurate experimental results. It is recommended to isolate the PCR reaction system preparation area from the PCR product purification area, and use a dedicated pipette to regularly clean each experimental area.
Schematic diagram of DNA library construction process
Starting material: 5ng-1μg of broken double stranded DNA, dissolved in EB (10 mM Tris HCl pH 8.0) or deionized water, DNA purity requirement: OD260/OD 280=1.8-2.0.
