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Invitrogen™ GLOBINclear™-Human Kit, for globin mRNA depletion
Description
Includes
20μL Capture Oligo Mix; 600μL Streptavidin Magnetic Beads; 1.5mL 2X Hybridization Buffer; 1.5mL Streptavidin Bead Buffer; 1.75 Nuclease-free Water; 200μL RNA Binding Beads; 5mL RNA Wash Solution Concentrate; 1mL Elution Buffer; 80μL RNA Bead Buffer; 4mL RNA Binding Buffer Concentrate; 100 1.5-ml Non-stick Tubes
Requires
Kit requires use of a magnetic stand. The Six-Tube Magnetic Stand (Cat. No. AM10055) can hold six separate 1.5mL conical microcentrifuge tubes or 2mL round-bottomed microtubes with a diameter of 8–10 mm.
The GLOBINclear-Human Kit is a rapid and robust treatment for RNA derived from human whole blood that uses a novel, non-enzymatic technology to rapidly deplete >95% of the alpha and beta globin mRNA from total RNA preparations.
Advantages of the GLOBINclear-Human Kit include:
• Improves the quality and accuracy of gene expression profiling by removing >95% of unwanted globin mRNA from whole blood total RNA to reveal biologically-relevant data
• Avoids harsh RNase H treatments that can degrade mRNA and alter expression profiles
• Produces a superior template for next-generation sequencing, RNA amplification reactions, cDNA synthesis, microarray analysis, and routine quantitative RT-PCR by using a non-enzymatic technology
Globin mRNA depletion is a vital preparatory step
Transcriptome expression profiling of whole blood samples is widely used to study physiological, pathogenic, and therapeutic responses of the transcriptome. However, up to 70% of the mRNA (by mass) in whole blood total RNA samples is composed of unwanted globin transcripts that affect the quality and accuracy of gene expression profiling in DNA microarray and sequencing technologies. Thus, globin mRNA depletion is a vital preparatory step to obtain accurate and biologically-relevant data during transcriptome analysis.
The GLOBINclear advantage
The GLOBINclear kit uses 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. First, globin mRNA is removed from whole blood RNA using a novel hybridization capture technology; then the remaining RNA is further purified with a rapid magnetic bead-based clean-up procedure.
As a result, GLOBINclear processed RNA provides more sensitive expression profile results than unprocessed whole blood RNA because the depletion procedure alleviates problems associated with the abundant globin mRNA found in whole blood RNA samples.
Furthermore, other globin mRNA depletion technologies rely on harsh RNase H enzymatic treatments that may compromise the expression profile by degrading mRNAs other than globin mRNA. Because the GLOBINclear kit uses a non-enzymatic technology, the resulting RNA is a superior template for next-generation sequencing, RNA amplification reactions, cDNA synthesis, microarray analysis, and routine quantitative RT-PCR.
Accessory product
The kit requires the use of a magnetic stand. The DynaMag-2 Magnetic Stand (Cat. No. 12321D) is recommended.
Order Info
Shipping Condition: Approved for shipment on Wet or Dry Ice
Specifications
Specifications
| Product Type | mRNA Depletion Kit |
| Content And 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) All components shipped on wet ice except for Capture Oligo Mix (dry ice). |
| Final Product Type | Total RNA (Globin mRNA-Depleted) |
| High-throughput Compatibility | High-throughput Compatible |
| Sample Type | Total RNA (Whole Blood) |
| Species | Human |
| Format | Microtube |
| Product Line | Ambion, GLOBINclear |
| For Use With (Application) | Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), Northern Blotting, cDNA Library Construction |
| Quantity | 20 Preps |
Frequently Asked Questions (FAQs)
To maximize removal of globin mRNA, follow the procedure exactly. Also pay attention to the most crucial parts of the procedure listed below:
-The GLOBINclear Kit procedure is optimized for depletion of globin mRNA from 10 µg or less total RNA from human, mouse, or rat whole blood. Using more than 10 µg RNA in the procedure will compromise removal of globin mRNA.
-Be sure to follow the temperature and time recommendations for the hybridizations in steps II.D.4 on page 9 and II.E.2 on page 10 of the manual (https://tools.thermofisher.com/content/sfs/manuals/fm_1980.pdf) for optimal depletion of globin mRNA.
The GLOBINclear Kit does not contain capture probes for ribosomal RNA (rRNA) sequences; however, after globin mRNA depletion with the kit, the percentage of ribosomal reads for the kit is only 1% of total reads (https://pubmed.ncbi.nlm.nih.gov/33308163/). This low percentage is irrelevant for most downstream applications.
For Research Use Only. Not for use in diagnostic procedures.