Introduction

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue and section samples
Applications	RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE tissue sample
Sample amount	6mg
Yield	20μg
Elution volume	≥20μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 60 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R414302	D414303
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	250
2ml Collection Tubes	50	250
Buffer DPS	60 ml	250 ml
Buffer FRL	15 ml	60 ml
Buffer RLC	15 ml	60 ml
Buffer RWC*	10 ml	50 ml
Buffer RW2*	20 ml	2 x 50 ml
Protease Dissolve Buffer	1.8 ml	10 ml
Proteinase K	24 mg	120 mg
RNase Free Water	10 ml	20 ml

Storage and Stability

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 their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Overview

• Purification and enrichment of intact urinary exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• Versatile urine input volumes
• No phenol extractions, Proteinase K treatment, nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Compatible with urine from most species
• Pure exosomes are purified and are free from any other RNA-binding proteins
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin
• The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services

Norgen’s Exosome Purification and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the subsequent isolation of exosomal RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified RNA is free from any protein-bound circulating RNA and of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Urine Exosome Purification and RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome Purification and RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome Purification and RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	11 mL
Maximum Urine Input	30 mL
Size of Exosomes Purified	40 nm – 150 nm
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35 – 40 minutes
Average Yields*	Variable depending on specimen

*Please check page 4 of the product insert for the average yields and the common RNA quantification methods

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 58400 (50 preps)	Cat. 58700 (25 preps)	Cat. 58800 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	20 mL	20 mL	20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	50	25	15
Collection Tubes	50	25	15
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

Introduction

Usages:
For selective enrichment of Salmonella.

Principle:
Peptone provide carbon and nitrogen sources to meet the needs of bacterial growth; lactose are fermentable sugars; selenite, sodium hydrogen inhibit Gram-positive bacteria and gram-negative enterobacteria most non-Salmonella; phosphate-buffered agent; L- cystine as a reducing agent.

Formulation(per liter):
Peptone: 5g
Lactose :4g
Sodium selenite: 4g
Disodium hydrogen phosphate: 10g
L- cystine: 0.01g
Final pH 7.0 ± 0.2

How to use:
1. Weigh 23g of the product , adding 1 L of distilled or deionized water , heated to boiling stirring until completely dissolved, dispensing flask, cooled to room temperature .
2. Pipette 10mL of pre-enrichment sample broth or 25mL and transferred species in the liquid sample flask in a sterile environment.
3.Place into incubator, cultured at 36 ± 1 for 18-24h.
4. Observe the results.

Quality control:
Quality control strains were inoculated ,and cultured at 36 ± 1 for 18-24h ,results show as follows:
strain name strain code growth feature
Salmonella typhi CMCC (B) 50071 good red, cloudy
Salmonella typhimurium CMCC (B) 50115 good red, cloudy
Escherichia coli ATCC25922 — remain unchanged

Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.

500g