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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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MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.
This method is suitable for small quantities of tissue (<100mg) and cells (<5 X106), and large quantities of tissue (up to 1g) and cells (<108), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.
Specifications
| Features | Specifications |
| Main Functions | RNA isolation solvent (substitution for Trizol/Qiazol reagent) |
| Applications | RT-PCR, Northern hybridization, poly (a) enrichment, etc. |
| Purification technology | Acid phenol guanidine isothiocyanate |
| Process method | Manual (centrifugation) |
| Sample type | Animal tissue, plant tissue, adherent cells, suspended cells, bacteria, trace fungi, etc. |
| Sample amount | Flexible |
| Elution volume | Variation with sample size |
| Time per run | Variation with sample size |
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.
Experiment Data
MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.
Introduction
A highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration. This kit can be easily used in manual and automated 96 or 384-well formats.
Specifications
| Features | Specifications |
| Main Functions | Recover 100-400μl DNA/ RNA from PCR products / enzymatic reaction solution / or crude DNA / RNA |
| Applications | Automatic sequencing, enzyme digestion, PCR and labeling |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Agarose Gel, PCR products, enzymatic reaction solution |
| Sample amount | 100-400µl |
| Recovery | 80% |
| Elution volume | ≥30μl |
The MagPure method contains magnetic particles in an optimized binding buffer to selectively bind DNA fragments 100bp and larger toparamagnetic beads. Excess primers, nucleotides, salts, and enzymes can be removed using a simple washing procedure.
Advantages
Kit Contents
| Contents | D500301 | D500302 | D500303 |
| Purification Times | 50 Preps | 500 Preps | 5000 Preps |
| Buffer AL | 10 ml | 60 ml | 550 ml |
| Buffer BD* | 5 ml | 25 ml | 2 x 100 ml |
| MagPure RNA Particles | 1.2 ml | 12 ml | 120 ml |
| RNase Free Water | 5 ml | 30 ml | 250 ml |
Storage and Stability
MagPure RNA Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
A highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration. This kit can be easily used in manual and automated 96 or 384-well formats.