Bis-propargyl-PEG9 is crosslinking reagent containing two propargyl groups. These functional groups enable the formation of triazole linkages with azide-bearing compounds with the presence of copper catalyst. The hydrophilicity of the molecule is improved because of the PEG chain. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bis-propargyl-PEG9 is crosslinking reagent containing two propargyl groups. These functional groups enable the formation of triazole linkages with azide-bearing compounds with the presence of copper catalyst. The hydrophilicity of the molecule is improved because of the PEG chain. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. 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 technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
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The Small RNA Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform. All molecular reagents including adaptors, primers, enzyme mixes and buffers are provided. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patent resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.
Workflow
Figure 1 / 4
Click for expanded view
| Kit Specifications | |
| Number of preps | 24 |
| Number of indices provided | 12 (for 24 preps, indexes 1-24 or 25-48) |
| Time required to complete 6 libraries | As little as 5 hours |
| Input RNA required | As little as 50 pg |
Storage Conditions and Product Stability
Some components require storage at -20°C, 4°C or room temperature. See individual components and box labels for storage conditions.
| Step | Component | Cat. 64600 (24 preps) |
|---|---|---|
| 3′ AdaptorLigation to Template RNA | 3′ Adaptor | 30 µL |
| 3′ Adaptor Ligation Master Mix | 320 µL | |
| T4 RNA Ligase 2 (Truncated) | 35 µL | |
| 5′ Adaptor Ligation | 5′ Adaptor | 30 µL |
| 5′ Adaptor Ligation Master Mix | 320 µL | |
| T4 RNA Ligase 1 | 35 µL | |
| cDNA Synthesis from Ligated RNA Product | Reverse Primer | 30 µL |
| cDNA Synthesis Master Mix | 220 µL | |
| TruScript ReverseTranscriptase | 35 µL | |
| PCR Amplification | 2x NGS PCR Master Mix | 1.32 µL |
| PCR Reverse Primer | 81 µL | |
| Forward Index Primer | Included in Small RNA Library Prep Forward Index Primers (# 64640 or # 64610) | |
| Size Selection | NGS MW Ladder | 50 µL |
| NGS Control Ladder | 50 µL | |
| Loading Dye | 300 µL | |
| Nuclease-Free Water | 1.25 µL |
| Component | Cat. 63500 (75 preps) |
|---|---|
| Buffer RL | 40 mL |
| Wash Solution A | 38 mL (Reconstituted to 128 mL) |
| Elution Solution A | 6 mL |
| Columns | 75 |
| Gel Filtration Columns | 24 |
| Collection Tubes | 75 |
| Elution Tubes | 75 |
| Product Insert | 1 |
Opentrons OT-2 Single-Channel Electronic Pipettes automate liquid handling and pipetting for most daily wetlab applications. Best for low to medium throughput workflows with high precision.
Opentrons OT-2 Single-Channel Electronic Pipettes automate liquid handling and pipetting for most daily wetlab applications. Best for low to medium throughput workflows with high precision.
