Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 1-10μl blood, <10mg tissue, urine, blood stain, seminal stain
Applications	PCR, southern bolt and virus detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissues, blood stain, urine, seminal stain and various forensic samples
Sample amount	Blood:1-100μl, Tissue:<10mg
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA).

Advantages

• Fast – several samples can be extracted in 20 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Contents	D312502	D312503
Purification Times	50 Preps	250 Preps
Buffer ATL	15 ml	60 ml
Buffer AL	15 ml	60 ml
Buffer GW1*	22 ml	66 ml
Buffer GW2*	20 ml	2 x 50 ml
Carrier RNA	310 μg	2 x 310 µg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	15 ml	60 ml
HiPure DNA Mini Columns I	50	2 x 125
2 ml Collection Tubes	100	5 x 100

Storage and Stability

Carrier RNA and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.

Experiment Data

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

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