Reagents - Microarray Labeling - RNA Extraction Kits to Prepare High Quality RNA for DNA Microarray Applications in Research, Life Sciences, Diagnostics and Pharmaceuticals

These kits are optimized for use with 10,000-2,000,000 cells as starting material, providing 2-25 µg of total RNA for microarray hybridizations and other life sciences applications. Use these kits in conjunction with Arrayit MiniAmp mRNA Amplification Kit and Indirect Amino Allyl Labeling Products.

Indirect Amino Allyl Fluorescent Labeling Kit
Universal Reference mRNA for Human, Mouse and Rat
MiniAmp mRNA Amplification Kit

Table of Contents

• Introduction
• Quality Control
• Product Description
• Kit Contents
• Technical Assistance
• Requirements
• Total RNA Extraction Protocol
• Ordering Information
• Warranty

Introduction
Congratulations on taking a big step towards improving the economies of scale, quality and speed of your genomics research. This booklet contains a complete set of protocols outlining the steps and principles needed to use Arrayit Micro Total RNA Extraction Kit.

Quality Control
Arrayit assures the performance of this product. The finest scientific research went into the development of this product. Rigorous quality control monitoring on a lot-by-lot basis guarantees that the ingredients conform to the highest industry standards.

Product Description:

• This kit is optimized to use with 10,000-2,000,000 cells as starting material and extract 2-25 µg total RNA for down stream experiments such as DNA microarrays.  To be used in conjunction with ArrayIt MiniAmp mRNA Amplification Kit and In-direct Amino Allyl Labeling Products.
• For preparation of total RNA in high yield and purity from a variety of tissue samples and cells
• Great for us with DNA microarrays
• Can be used to isolate RNA from tissues and cells, including plant, animal and yeast cells and protozoa.
• Yields total RNA suitable for direct use in Northern blot hybridizations and in vitro translation, or as starting material for purification of mRNA for cDNA synthesis.

Kit Contents

Usage
This kit is optimized to use with 10,000-2,000,000 cells as starting material and extract 2-25 µg total RNA for down stream experiments such as DNA microarrays. Samples may include whole blood, cells or tissue.

Requirements:
Items needed but not supplied

• 1.5 ml centrifuge tubes
• 100% ethanol
• Ice for incubations
• 1 ml disposable syringes with 20 gauge needles

Equipment needed

• Micropipettes
• Vortex mixer
• Microcentrifuge

Total RNA Extraction Protocol from whole blood/cells/tissue

• Spin all tubes 5-10 second before use.
• Add 2 ml of 100% ethanol to RNA Wash Buffer Concentrate before starting
• NOTE: this system is compatible with any type of anti-coagulant, citrate, EDTA, Heparin...
• All 200 µl of whole blood, or cells or tissue to 2 ml of RBC Lysis Buffer and Incubate on ice for 15 minutes. NOTE: this step can be skipped when working with cells in culture.
• Collect cell pellet by gentle centrifugation at 500 x g for 5 minutes.
• Remove and discard the supernatant.
• Add 100 µl of RNA Extraction Buffer to the cell pellet and resuspend gently by pipetting.
• Homogenize the sample using a 1 ml disposable syringe with 20-gauge needle
  (5-8 times).
• Spin 1 sec in a microcentrifuge by briefly “pulsing” at low speed. If a cell pellet forms, repeat homogenization.  If not, proceed with the extraction.
• Incubate on ice for 20 minutes.
• Add 100 µl of 100% ethanol, mix briefly and incubate on ice for 10 minutes.
• Load the entire contents onto an RNA Micro Column and place the column in a 2 ml wash tube.
• Centrifuge for 1 minute at full speed in a microfuge (>10,000 rpm), discard the flow through and reuse wash tube.
• Add 100 µl of RNA Wash Buffer to the column and centrifuge at full speed in a microfuge (>10,000 rpm) for 1 minute.
• Add another 100 µl of RNA Wash Buffer Concentrate to the column and centrifuge at full speed in a microfuge (>10,000 rpm) for 1 minute (NOTE: longer centrifugation times may be necessary to remove residual wash buffer.
• Discard the 2 ml wash tube and carefully place the column in an elution tube.
• Add 40 µl of nuclease-free water pre-warmed to 60°C into the center of the column
• Incubate 2 minutes and centrifuge at full speed in a microfuge (>10,000 rpm) for 1 minute.
• Re-load the filtrate onto the column, incubate 2 minutes and centrifuge at full speed in a microfuge (>10,000 rpm) for 1 minute.
• Discard the column and use the eluted total RNA for downstream applications.