GLOBINclear™-Human Kit, for globin mRNA depletion
GLOBINclear™-Human Kit, for globin mRNA depletion
Invitrogen™

GLOBINclear™-Human Kit, for globin mRNA depletion

The GLOBINclear-Human Kit is a rapid and robust treatment for RNA derived from human whole blood that uses a novel,Read more
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Catalog number AM1980
Price (USD)
668.00
Each
Add to cart
Price (USD)
668.00
Each
Add to cart
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.

Alternative formats
The GLOBINclear-Human Kit is also available in an automation-friendly 96-well format for high throughput screening. For more information, please contact Technical Support.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypemRNA Depletion Kit
Final Product TypeTotal RNA (Globin mRNA-Depleted)
High-throughput CompatibilityHigh-throughput Compatible
Sample TypeTotal RNA (Whole Blood)
SpeciesHuman
FormatMicrotube
Product LineAmbion™, GLOBINclear™
For Use With (Application)Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), Northern Blotting, cDNA Library Construction
Quantity20 Preps
Unit SizeEach
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)

All components shipped on wet ice except for Capture Oligo Mix (dry ice).

Frequently asked questions (FAQs)

I used the GLOBINclear Kit to deplete globin mRNA from my total RNA sample, but the depletion was not very efficient. What should I do?

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.

Does the GLOBINclear Kit deplete ribosomal RNA (rRNA) in addition to 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.