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 (with DNase)
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, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE tissue sample
Sample amount	6mg
Yield	20μg
Elution volume	≥10μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

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
• Efficient DNA removal – unique method to effectively remove genomic DNA

Kit Contents

Contents	R414402	D414403
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
DNase I	600 µl	5 x 600 µl
DNase Booster Buffer	1.5 ml	6 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. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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.

Experiment Data

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

• Adenovirus Purification from any input – cell fraction or media fraction
• Rapid purification within 2 to 4.5 hours
• No specialized equipment needed (ultracentrifuge not required)
• Purify adenovirus cell culture supernatant from 1 mL to 33.5 mL input per prep
• Purify adenovirus cell pellet from 1 mL of input per prep
• Up to 25X sample concentration
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Adenoviral vectors are useful tools for both in vitro and in vivo gene transfer, and oncolytic viruses based on adenovirus are highly promising for cancer treatment. Norgen’s Adenovirus Purification Kit provides a fast and simple procedure for concentrating and purifying adenoviral vectors from cell lysate and cell culture media. Purification is based on spin column chromatography using Norgen’s proprietary resin as an ion exchanger.  Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. Adenoviral vectors purified in this manner are highly active for use in transduction experiments.

Norgen’s Adenovirus Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. 

Details

Supporting Data

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Kit Specifications
Resin Binding Capacity (total per kit)	At least 3×1011 adenovirus particles as determined by qPCR At least 3×108 transducing units as determined by transduction assay
Input type	Cells, media
Input volume (Supernatant)	1 – 33.5 mL SN per prep (500 mL SN in total)
Input Volume (Cell pellet)	1 mL cell pellet per prep (15 mL in total)
Minimum elution volume	1.3 mL per prep
Time to complete purification	2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction	Yes

Storage Conditions and Product Stability

HL-SAN Nuclease should be stored at -20°C upon arrival. Elution Buffer P should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 67600 (15 preps)
Lysis Buffer S	5.5 mL
HL-SAN Nuclease	102 µL
Binding Buffer A	20 mL
Purification Solution C	60 mL
Purification Solution D	130 mL
Wash Solution C	2 x 130 mL
Slurry E	12.5 mL
Elution Buffer P (store at 4°C)	66 mL
Protein Neutralizer	4 mL
Elution tubes (1.7 mL)	50
Product Insert	1