Overview
- Optimized for low input RNA, especially from bodily fluids such as plasma or serum at 1ng of RNA
- Simple and quick workflow: library could be prepared in less than 5 hours
- No gel purification for selected types of samples
- Protocol optimized for RNA isolated from different types of input, including liquid biopsies (blood, plasma, serum and urine)
- Complements Norgen’s Best-in-Class Total RNA (including microRNA) Purification Technology
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
Details
Supporting Data
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 |
