GLOBINclear™ Kit, mouse/rat, for globin mRNA depletion

Catalog number: AM1981

Invitrogen™  Related applications: RNA Extraction

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Description

The Ambion® GLOBINclear™-Mouse/Rat Kit employs a novel, non-enzymatic technology that rapidly depletes >95% of the alpha and beta globin mRNA from total RNA preparations derived from whole blood. The kit includes sufficient reagents for 20 reactions.

• Remove >95% of unwanted globin mRNA from whole blood total RNA
• Detect up to 50% more genes—analyze previously undetected genes
• Achieve greater representation of the genome, low 3'/5' ratios
• Avoid RNase H treatments that can degrade mRNA and alter expression profiles

It is widely known that expression array data generated from whole blood total RNA samples have reduced detection sensitivity compared to data from fractionated blood samples. Examination of whole blood total RNA reveals a dominant band in the 600–700 base region. This band represents globin mRNA, which is expressed at high levels in red blood cells (RBCs) and reticulocytes (RBC precursors). Up to 70% of the mRNA (by mass) in whole blood total RNA are globin transcripts. Array analysis using total RNA containing a high percentage of globin mRNA has been shown to:

• Decrease present calls
• Decrease call concordance
• Increase signal variation

The GLOBINclear™ kit employs a novel hybridization technology that takes advantage of the strength of biotin/streptavidin binding, the specificity of nucleic acid hybridization, and the convenience of magnetic separations. There are no harsh RNase H enzymatic treatments that may compromise the expression profile by degrading mRNAs other than globin mRNA. The resulting mRNA is a superior template for synthesizing labeled cDNA for array analysis and is ideal for quantitative RT-PCR. The enrichment delivers a significant increase in Percent Present calls, decreased sample-to-sample variability, and low 3'/5' ratios.

Accessory Product:
The kit requires the use of a magnetic stand. The 6 Tube Magnetic Stand (SKU# AM10055) can hold six separate 1.5 mL conical microcentrifuge tubes or 2 mL round-bottomed microtubes with a diameter of 8–10 mm.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Sample Type (General): RNA, Blood
High Throughput Compatibility: High Throughput-Compatible
Downstream Application: Reverse Transcriptase PCR (RT-PCR), Northern Blotting, Real-Time Quantitative PCR (qPCR), Microarray Analysis, cDNA Library Construction
Scale: Mini
Format: 6 microtubes
Product Size: 20 preps
Final Product: Total RNA (Globin mRNA-Depleted)
Yield (up to): 5 µg
Purification Time: 1.5 hrs
Number of Reactions: 20 Reactions
Isolation Technology: Magnetic Bead
Sample Type (Specific): Total RNA (Whole Blood)
Starting Material (Amount): 1 - 10 µg of mouse or rat total RNA from whole blood

Contents & storage

• 20 µl Capture Oligo Mix (-20°C)
• 600 µl Streptavidin Magnetic Beads (4°C—do not freeze)
• 1.5 ml 2X Hybridization Buffer (room temperature)
• 1.5 ml Streptavidin Bead Buffer (room temperature)
• 1.75 Nuclease-free Water (-20°C, 4°C, or room temperature)
• 200 µl RNA Binding Beads (4°C—do not freeze)
• 5 ml RNA Wash Solution Concentrate (4°C or room temperature)
• 1 ml Elution Buffer (4°C or room temperature)
• 80 µl RNA Bead Buffer (room temperature)
• 4 ml RNA Binding Buffer Concentrate (room temperature)
• 100 1.5-ml Non-stick Tubes (room temperature)

Documents

Manuals & protocols