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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.
Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.
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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.
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SKU: CB0215
Permagen’s Thermal Combo Block 1.5 mL, 0.2 mL PCR was designed as a temperature control device that can be used manually or on a liquid handling robot
This product is suitable for rapid extraction of RNA from low RNA yield somples such as tissue (＜10mg), cells, bone marrow, fresh blood, and other clinical samples. RNA can be used directly for RT-PCR, Real time PCR, NGS, Viral RNA detection and so on.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from tissue, cell, whole blood |
| Applications | RT-PCR, cDNA synthesis, second generation sequencing |
| Purification method | Polydisperse magnetic beads |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Adaptive instrument | Nucleic acid extractor, pipetting workstation |
| Sample type | Tissues, cells, lymphocytes and other clinical sample |
| Sample amount | Cells grown in suspension: ≤3 x 106Animal tissue: ≤10mgPlant tissue: ≤30mgWhole blood: 0.5~1.0 ml fresh blood or bone marrow and fresh blood mixture |
Kit Contents
| Contents | IVD3020B-96 | IVD3020B |
| Purification Times | 96 Preps | 200 Preps |
| MagPure Particles N | 2.5 ml | 5 ml |
| DNase I | 2 x 600 µl | 4 x 600 µl |
| DNase Buffer C | 60 ml | 120 ml |
| Buffer RLC | 60 ml | 120 ml |
| Buffer MCB* | 60 ml | 150 ml |
| Buffer GW1* | 44 ml | 110 ml |
| Buffer MW2* | 50 ml | 100 ml |
| RNase Free Water | 20 ml | 60 ml |
Storage and Stability
DNase I should be shipped with ice pack and stored at -20°C after arrival. MagPure Particles N should be stored at 2–8°C for long time storage. The remaining kit components can be stored at room temperature(15–25°C) for up to18 months under these conditions.
This product is suitable for rapid extraction of RNA from low RNA yield somples such as tissue (＜10mg), cells, bone marrow, fresh blood, and other clinical samples. RNA can be used directly for RT-PCR, Real time PCR, NGS, Viral RNA detection and so on.