Universal 23s PCR Amplification Kit (For targeted next-generation sequencing)

Propargyl-PEG8-amine is a heterobifunctional reagent consisting of a propargyl group and an amine group. The amine group can form amide bonds with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can form triazole linkage with azides in copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG8-amine is a heterobifunctional reagent consisting of a propargyl group and an amine group. The amine group can form amide bonds with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can form triazole linkage with azides in copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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

Previous

Next

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