These kits provide a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. These kits are supplied with an RBC (red blood cell) Lysis Buffer for selective removal of red blood cells and fractionation of leukocytes by centrifugation. Isolation of leukocyte RNA results in improved expression profiling and other downstream applications by removing the masking effects of some RNAs which are very abundant in whole blood, such as globin mRNAs. These kits are able to isolate total leukocyte RNA, including both large mRNA and all small RNA species containing microRNA (miRNA) and small silencing RNA (siRNA). The purified RNA is of the highest quality and can be used in a number of downstream applications.
Leukocyte RNA Purification Kit (Spin Column)
This kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Allowable blood input ranges from 10 μL to 2 mL or 3 x 106 Leukocytes
Leukocyte RNA Purification Plus Kit (Plus)
Norgen’s Leukocyte RNA Purification Plus Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from up to 3 mL of mammalian blood samples. Complete 10 purifications in as little as 40 minutes.
Leukocyte RNA Purification 96-Well Kit (High Throughput)
Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Norgen’s kit allows for the isolation of total leukocyte RNA, including all small RNA species. The purified RNA is of the highest quality and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, northern blotting, RNase protection and primer extension, and expression array assays. Allowable blood input ranges from 10 μL to 1 mL or 1.5 x 106 Leukocytes.
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| Kit Specifications | |
| Maximum Binding Capacity Per Well | 50 μg |
| Maximum Loading Volume Per Well | 500 μL |
| Size of RNA Purified | All sizes, including small RNA < 200 nt |
| Maximum Blood Input* | 1 mL or 1.5 x 106 Leukocytes |
| Standard Blood Input | 150 μL |
| Minimum Blood Input | 10 μL |
| Average Yield: 500 μL human blood | 1.5 μg |
| Time to Complete 10 Purification | 40 minutes |
*Additional RBC Lysis Solution is required for input Volumes >150 µL
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The RBC Lysis Buffer should be stored at 4°C upon arrival. These reagents should remain stable for at least 1 year in their unopened containers.
| Component | Cat. 21200 (50 preps) | Cat. 21250 (50 preps) | Cat. 37800 (2 x 96 preps) |
|---|---|---|---|
| RBC Lysis Buffer | 2 x 100 mL | 2 x 1 L | 2 x 90 mL |
| Buffer RL | 30 mL | 30 mL | – |
| Binding Solution | – | – | 2 x 40 mL |
| Wash Solution A | 38 mL | 38 mL | – |
| Wash Solution | – | – | 2 x 30 mL |
| Elution Solution A | 6 mL | 6 mL | – |
| Elution Solution | – | – | 2 x 20 mL |
| Enzyme Incubation Buffer | – | 6 mL | 1 |
| DNase I | – | 1 Vial | – |
| Mini Spin Columns | 50 | – | – |
| Lysate Homogenization Column | – | 50 | – |
| Single Cell RNA Column | – | 50 | – |
| RBC Lysis 96-Well Plate | – | – | 2 |
| 96-Well Filter Plate | – | – | 2 |
| Adhesive Tape | – | – | 4 |
| Collection Tubes | 50 | 100 | – |
| 96-Well Collection Plate | – | – | 2 |
| Elution Tubes (1.7 mL) | 50 | 50 | – |
| 96-Well Elution Plate | – | – | 2 |
| Product Insert | 1 | 1 | 1 |
Bis-propargyl-PEG18 has two propargyl groups at both ends. These propargyl groups reacts with azide compounds or biomolecules to form stable triazole linkages under the catalyzation of copper. The hydrophilic PEG units enhance the solubility of the molecule in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bis-propargyl-PEG18 has two propargyl groups at both ends. These propargyl groups reacts with azide compounds or biomolecules to form stable triazole linkages under the catalyzation of copper. The hydrophilic PEG units enhance the solubility of the molecule in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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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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