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
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| 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 |
Propargyl-PEG4-CH2CO2H is a linker consisting of a propargyl group with a carboxylic acid group. The carboxylic acid can react with primary amine groups in the presence of activators such as HATU or EDC. The propargyl group can react with azide compounds in Click Chemistry; copper catalyst will be needed. The PEG units help the molecule to have better solubility in aqueous solution. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG4-CH2CO2H is a linker consisting of a propargyl group with a carboxylic acid group. The carboxylic acid can react with primary amine groups in the presence of activators such as HATU or EDC. The propargyl group can react with azide compounds in Click Chemistry; copper catalyst will be needed. The PEG units help the molecule to have better solubility in aqueous solution. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
West Nile Virus (WNV) belongs to the RNA virus family of Flaviviridae. It is known to spread primarily through arthropod vectors such as mosquitoes. The virus infects mainly birds but is also reported to infect humans as well as pets such as cats and dogs. In humans, the majority of the infections caused by WNV are asymptomatic. However, some individuals (the majority of the confirmed cases) could enter a second, febrile stage with flu-like symptoms – commonly known as West Nile Fever. In a more serious stage, the disease becomes neuroinvasive, causing meningitis or encephalitis. Such severe conditions could lead to mortality. As the symptoms of WNV are very similar to other common diseases but can be fatal at a severe stage, it is important to distinguish the virus early in the diagnosis, particularly at the molecular level.
WNV TaqMan RT-PCR Kit, 100 reactions
WNV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
Figure 1 / 3
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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM44250 (100 preps) | Cat. TM44210 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| WNV Primer & Probe Mix | 280 μL | 280 μL |
| WNV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
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