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

Overview

• Fractionate leukocytes from whole blood in minutes with provided RBC Lysis Buffer
• Isolate total RNA, including microRNA, without phenol
• Rapid and convenient spin-column format and 96-well plate for high throughput applications
• Purified RNA is ready for any downstream application including RT-PCR, qRT-PCR, NGS, arrays and more. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. These kits are supplied with an RBC (red blood cell) Lysis Buffer for selective removal of red blood cells and fractionation of leukocytes by centrifugation. Isolation of leukocyte RNA results in improved expression profiling and other downstream applications by removing the masking effects of some RNAs which are very abundant in whole blood, such as globin mRNAs. These kits are able to isolate total leukocyte RNA, including both large mRNA and all small RNA species containing microRNA (miRNA) and small silencing RNA (siRNA). The purified RNA is of the highest quality and can be used in a number of downstream applications.

Leukocyte RNA Purification Kit (Spin Column)

This kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Allowable blood input ranges from 10 μL to 2 mL or 3 x 10⁶ Leukocytes

Leukocyte RNA Purification Plus Kit (Plus)

Norgen’s Leukocyte RNA Purification Plus Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from up to 3 mL of mammalian blood samples. Complete 10 purifications in as little as 40 minutes.

Leukocyte RNA Purification 96-Well Kit (High Throughput)

Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Norgen’s kit allows for the isolation of total leukocyte RNA, including all small RNA species. The purified RNA is of the highest quality and 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. Allowable blood input ranges from 10 μL to 1 mL or 1.5 x 10⁶ Leukocytes.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 μg RNA
Maximum Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA <200 nt
Minimum Blood Input	10 μL
Maximum Blood Input	2 mL or 3 x 106 leukocytes
Time to Complete 10 Purifications	40 minutes
Input Type	Blood from Human, Hamster, Mouse, Rabbit, and Rat
Average Yield Human Blood (500 μL) Hamster Blood (100 μL)	1.5 μg 2.5 μg

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The RBC Lysis Buffer should be stored at 4°C upon arrival.

Component	Cat. 21200 (50 preps)	Cat. 21250 (50 preps)	Cat. 37800 (2 x 96 preps)
RBC Lysis Buffer	2 x 100 mL	2 x 1 L	2 x 90 mL
Buffer RL	30 mL	30 mL	–
Binding Solution	–	–	2 x 40 mL
Wash Solution A	38 mL	38 mL	–
Wash Solution	–	–	2 x 30 mL
Elution Solution A	6 mL	6 mL	–
Elution Solution	–	–	2 x 20 mL
Enzyme Incubation Buffer	–	6 mL	1
DNase I	–	1 Vial	–
Mini Spin Columns	50	–	–
Lysate Homogenization Column	–	50	–
Single Cell RNA Column	–	50	–
RBC Lysis 96-Well Plate	–	–	2
96-Well Filter Plate	–	–	2
Adhesive Tape	–	–	4
Collection Tubes	50	100	–
96-Well Collection Plate	–	–	2
Elution Tubes (1.7 mL)	50	50	–
96-Well Elution Plate	–	–	2
Product Insert	1	1	1

Introduction

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from bacteria, yeast cells
Applications	RT-PCR, Northern blot, 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	Bacteria, yeast cells
Sample amount	Bacteria: <10^9；Yeast cells：<2 x10^7
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

Advantages

• Fast – several samples can be extracted in 30 minutes
• High purity – the purified RNA can be directly used in various downstream applications
• High recovery – RNA can be recovered at the level of PG
• Good repeatability – silica gel column purification technology can obtain ideal results every time
• Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
• Sufficient components – the kit contains carried wall breaking enzymes and glass beads

Kit Contents

Contents	R418202	R418203
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1-0.6mm)	30 g	150 g
DNase I	600 μl	5 x 600 μl
DNase Buffer	6 ml	30 ml
Protease Dissolve Buffer	1.8 ml	15 ml
Buffer ATL	50 ml	200 ml
Buffer PHC	50 ml	200 ml
Buffer GXP2*	20 ml	100 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Buffer PHC 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, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.