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 Binding Capacity Per Well	50 μg
Maximum Loading Volume Per Well	500 μL
Size of RNA Purified	All sizes, including small RNA < 200 nt
Maximum Blood Input*	1 mL or 1.5 x 106 Leukocytes
Standard Blood Input	150 μL
Minimum Blood Input	10 μL
Average Yield: 500 μL human blood	1.5 μg
Time to Complete 10 Purification	40 minutes

*Additional RBC Lysis Solution is required for input Volumes >150 µL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature.  The RBC Lysis Buffer should be stored at 4°C upon arrival.  These reagents should remain stable for at least 1 year in their unopened containers.

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

Overview

• Sequentially purify total RNA (and miRNA), DNA and proteins from a single sample
• No sample splitting or need to use phenol or precipitation methods
• Purify RNA/DNA/Protein from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

RNA/DNA/Protein Purification Plus Kit

This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.

RNA/DNA/Protein Purification Plus Micro Kit

This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.

RNA/DNA/Protein Purification 96-Well Plus Kit

The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.

Details

Supporting Data

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Kit Specifications
Binding Capacity Per Well	50 μg for RNA 20 μg for DNA 150 μg for protein
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Size of DNA Purified	≥ 30 kb
Time to Complete 10 Purifications	35 minutes
Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodBacteriaYeastFungiPlant Tissues	1 x 106 cells10 mg100 µL1 x 109 cells1 x 108 cells40 mg40 mg
Average Yield* Liver (5 mg)	12.5 μg RNA 2 μg DNa  55 μg Protein

* average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.

Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT).

Component	Cat. 47700 (50 preps)	Cat. 51600 (50 preps)	Cat. 51700 (96 preps)
Buffer SKP	40 mL	40 mL	–
Lysis Buffer Q	–	–	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL	20 mL
Elution Buffer F	15 mL	15 mL	2 x 15 mL
Wash Solution C	30 mL	30 mL	60 mL
Binding Buffer A	8 mL	8 mL	8 mL
Elution Buffer C	8 mL	8 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Protein Loading Dye	2 mL	2 mL	3 x 2 mL
gDNA Purification Columns	50	–	–
gDNA Purification Micro Columns	–	50	–
gDNA Purification 96-Well Plate	–	–	1
RNA/Protein Purification Columns	50	–	–
RNA/Protein Purification Micro Columns	–	50	–
RNA/Protein Purification 96-Well Plate	–	–	1
Collection Tubes	150	150	–
Collection Plate	–	–	5
Elution Tubes (1.7 mL)	150	150	–
Elution Plate	–	–	3
Lysis Preparation Plate	–	–	2
Adhesive Tape	–	–	4
Product Insert	1	1	1

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 whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 200μl whole blood
Applications	PCR, southern bolt and virus detection, etc
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	200μl
Elution volume	≥50μl
Time per run	≤60 minutes

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
• Fast – polydisperse magnetic beads, fast magnetic response and short extraction time
• High purity – the obtained DNA can be directly used for second-generation sequencing, PCR based detection, gene bank, etc.
• Automatic – saving time andlabor and safer

Kit Contents

Contents	D631101	D631102	D631103
Purification Times	48	96	480
MagPure Particles	1.2 ml	2.5 ml	11 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
Buffer AL	15 ml	30 ml	120 ml
Buffer BD	5 ml	10 ml	50 ml
Buffer GW1	22 ml	53 ml	220 ml
Elution Buffer	15 ml	30 ml	100 ml

Storage and Stability

Proteinase K, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage(up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry 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.

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 whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.