Product Introduction
Aiming at Illumina high-throughput sequencing platform, this kit provides a convenient and universal DNA library construction scheme in one tube. It combines fragmentation, end repair and A-Tailing into one step, greatly shortening the time of library construction and reducing the error caused by tedious steps. After fragmentation and end preparation, the product can be directly ligated with adapter without additional purification, and the subsequent procedure is the same as that of #K001 Fast DNA Library Prep Kit. Complete library quantification can be performed by dsDNA fluorescent dye method (e.g., Thermo Qubit Flex Fluorometer) or absolute quantification PCR after diluting the library to an appropriate concentration.
Application
• NGS library preparation for Illumina platform
Features
1 Wide sample compatibility
2 High efficiency in library construction
3 Compatible with PCR-free workflow
4 Complete Fragmentation and End Repair simultaneously
Experimental Cases
1. High conversion rate of library.
Figure 1 Library conversion rate when DNA input was 60ng.
2. Whether before or after amplification, compared with other manufacturers, the amplification is very superior.
Figure 2 The concentration of unamplified library obtained when the DNA sample was 100ng and 1ng, and the end repair and adapter ligation were performed with the library construction kits of different manufacturers.
Figure 3 The total amount of library obtained after amplified with the same cycles when the DNA sample was 100ng, 60ng, and 20ng, and the end repair and adapter ligation were performed with the library construction kits of different manufacturers.
3. The time of fragmentation is flexible and controllable.
Figure 4 The input DNA was 500ng, and the fragmentation time was 8min, 12min, 15min, 18min, respectively. Image obtained by fragment size analysis and processing with Agilent Bioanalyzer 2100.
4. Strong product stability. The fragmentation effect is equally significant whether compared with the same batch or across batches.
Figure 5 Fragmentation effect between the same batch for 6min, 10min, 15min and 20min respectively.
Figure 6 Fragmentation effects between four different batches for 6min, 10min, 15min and 20min respectively.
Components
Component | K004-A (24 rxns) | K004-B (96 rxns) |
FEP Buffer | 120 μl | 480 μl |
FEP Enzyme Mix | 240 μl | 2×480 μl |
Fast DNA Ligase | 120 μl | 2×240 μl |
Fast Ligation Buffer | 600 μl | 4×600 μl |
2× HIFI Library PCR Master Mix | 600 μl | 4×600 μl |
Primer Mix | 120 μl | 480 μl |
Neutralization Buffer | 120 μl | 480 μl |
Storage Condition
All reagents should be stored at -20°C.