Description
SPLIT RNA Extraction Kit
The SPLIT RNA Extraction Kit enables a fast and highly efficient extraction of RNA that is free of genomic DNA contamination. The RNA can be recovered as total RNA or split into two fractions, large RNA and small RNA, facilitating the analysis of e.g., mRNA and miRNA from the same sample. Thus the RNA obtained is ideal for seamlessly preparing libraries for Next Generation Sequencing of total RNA or its large and small fractions or any other demanding downstream application. The SPLIT RNA Extraction Kit for Blood enables concomitant depletion of globin mRNAs from human blood samples in low volumes (50 – 250 µl).
SPLIT Rapid Viral RNA/DNA Extraction Kit is available for fast and easy extraction of viral RNA from fluid samples (e.g. for SARS-CoV-2 analysis).
High Quality, High Yield
RNA extracted with the SPLIT RNA Extraction Kit has a high RIN quality score for all types of samples. A RIN of 10 and a 28S / 18S rRNA ratio of 2.7 can be obtained from cell culture. Extractions from tissue samples usually result in RNA with a RIN of 8.0 – 9.5.
Small RNA and Large RNA Fractions
The SPLIT kit can be used for the extraction of either total RNA (< 17 nt to > 10,000 nt) or for the isolation of the large RNA fraction (cut-off at ~ 150 nt), with the option to obtain the small RNA fraction separately (Figure 1).
Rapid Turnaround
Efficient miRNA Recovery
Efficient recovery of siRNA and miRNA down to 17 nt in the total RNA or in the small RNA fraction has been shown in spike-in experiments with small RNA markers (Figure 2).
Figure 1. Agarose gel analysis of RNA samples extracted with the SPLIT kit or by a TRIzol / isopropanol precipitation method. In the TRIzol extracted sample genomic DNA is visible as a slot-retained band, whereas RNA obtained with the SPLIT kit is free from detectable genomic DNA contamination.
Free From Genomic DNA Contamination
Due to its highly optimized, phenol-extraction based protocol, the genomic DNA (gDNA) content in the extracted RNA sample is negligible compared to conventional methods (Figure 1).
No DNase Treatment, No RNA Degradation
The SPLIT protocol does not require DNase treatment which is often used for the removal of genomic DNA in the sample and can be a reason for degradation of RNA.
No gDNA Removal Column, No RNA Size Bias
The SPLIT workflow does not require the use of gDNA removal columns. Their function is based on size exclusion which can impose a size bias on the extracted RNA as well.
Figure 2. Separation of SPLIT RNA samples on a polyacrylamide gel, demonstrating the splitting of large and small RNA at a threshold of ~150 nt. The total RNA sample comprising small and large RNA is shown as comparison. The homogenate was spiked with a miRNA marker to assess efficient miRNA recovery.
RNA Extraction from Blood
The SPLIT RNA Extraction Kit for Blood includes an additional step which enables efficient depletion of predominant globin mRNA from human blood samples. This results in increased gene detection (Figure 3).
Figure 3. A) The SPLIT for Blood protocol depletes >95 % of globin mRNA species from the extracted RNA samples. RNA was extracted from fresh human blood using the SPLIT (left side bar) and SPLIT for Blood protocol (right side bar), respectively. RNA-Seq libraries were prepared with Lexogen’s QuantSeq 3’ mRNA-Seq (FWD) kit and sequenced reads were mapped to the human reference genome and the percentage of reads mapping to globins was calculated. B) Increased gene detection in human blood QuantSeq libraries using SPLIT RNA Extraction Kit for Blood. The number of discovered genes was calculated from CPM normalized read counts (threshold >0.5 CPM).
Viral RNA Extraction
Workflow
The SPLIT RNA Extraction Kit contains reagents for the isolation of total RNA or the large RNA fraction from 48 samples, or small and large RNA fractions from 24 samples.
For viewing the whole workflow on page please click here
Featured Publications
FAQ
Frequently Asked Questions
Please find a list of the most frequently asked questions below. If you cannot find the answer to your question here or want to know more about our products, please contact support@lexogen.com.
The SPLIT RNA Extraction Kit for Blood protocol is verified for use with 50 – 250 µl of fresh human blood collected in EDTA containing blood collection tubes. Please contact support@lexogen.com if you wish to use this protocol with blood from other species, variable input blood volumes, or with blood exposed to other collection methods or storage conditions.
The protocol is verified for use with 50 – 250 µl of fresh human blood collected in EDTA containing blood collection tubes. Please contact support@lexogen.com if you wish to use this protocol with blood from other species, variable input blood volumes, or with blood exposed to other collection methods or storage conditions.
Troubleshooting Guide
Problem | Likely cause | Comments and suggestions |
No or poor phase separation | Cooled phase-lock gel or insufficient centrifugation |
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Incorrect cut-off | Volume of isopropanol addition |
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Degraded RNA | RNA source |
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RNase contamination |
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Low absorption ratios (i.e., peak at 230 nm) | Contamination with organic solvents, salts, or metal ions |
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gDNA contamination | Contamination with organic phase |
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Low or no RNA yield | RNA remains on spin column |
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Spin column was overloaded |
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Problem in downstream application | Salt or ethanol carry-over during elution |
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Downloads
SPLIT RNA Extraction Kit
User Guide – update 10.01.2020
Send us your publication & get the RNA T-shirt!
Application Note – SPLIT RNA Extraction Kit
Product Flyer – SPLIT RNA Extraction Kit for Blood
SPLIT Supplementary Protocol: Purification of RNA from FFPE Samples – upload 18.07.2018
Material Safety Datasheets
MSDS Information can be found in the Documents page.
If you need more information about our products, please contact us through support@lexogen.com or directly under +43 1 345 1212-41.
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