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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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Description
AccuBand™ 100 bp+3K DNA Ladder II is composed of 14 individual DNA fragments, presenting 3k, 2k, 1.5k, 1.2k, 1k, 900, 800, 700 600, 500, 400, 300, 200 and 100 bp sharp bands respectively. This product contains 2 enhanced bands (1k and 500 bp) for easy band identification. AccuBand™ 100 bp+3K DNA Ladder II is ready-to-use, containing loading buffer with dual color tracking dyes (orange G and Xylene cyanol FF). AccuBand™ 100 bp+3K DNA Ladder II provides a sufficient amount of DNA for clear observation of all DNA bands ranging from 100 bp to 3 kb, either in agarose gel or in polyacrylamide gel electrophoresis.
Features
Source
Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
Range
100 ~ 3,000 bp
Concentration
54.4 µg/ 500 µl
Recommended loading volume
5 µl/ well
Storage
Room temperature for 6 months
4°C for 12 months
-20°C for 36 months
Clone | IHC655 |
Source | Mouse Monoclonal |
Positive Control | Prostate, Prostate Adenocarcinoma |
Dilution Range | 1:200 |
Prostatic Specific Acid Phosphatase (PSAP) is a prostatic enzyme found in the glandular epithelium of the prostate. PSAP levels are elevated in hyperplastic prostate and prostate carcinoma, with the highest levels being detected in metastasized prostate cancer. Moderate overexpression of PSAP is also characteristic of diseases of the bone (such as Paget’s disease or hyperparathyroidism), diseases of blood cells (such as sickle-cell disease), multiple myeloma, or lysosomal storage diseases (such as Gaucher’s disease). PSAP is considered more sensitive, yet less specific, than PSA, however Anti-PSAP can act as a useful complement to Anti-PSA under suitable clinical contexts.
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