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SPLIT RNA Extraction Kit

  • High-quality RNA for demanding downstream applications
  • Total RNA or split into small and large RNA fractions
  • Special versions for blood or virus RNA extraction available

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Description

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 (> 150 nt) and small RNA (< 150 nt), 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.

With the content of the kit total RNA or the large RNA fraction can be extracted from 48 samples. If the small RNA fraction should be purified in addition to the large RNA fraction, then 24 samples can be extracted.

The SPLIT RNA Extraction Kit for Blood additionally includes the Blood Lysis Buffer for lysis of red blood cells prior to sample homogenization of the conventional SPLIT RNA Extraction Kit. This additional step enables efficient depletion of globin mRNA from the resulting RNA sample from as little as 50 to 250 µl of human blood.

Kit Content:

  • Buffers and Purification Columns
  • Phase-Lock Gel Columns

008 (SPLIT RNA Extraction Kit),

099 (SPLIT RNA Extraction Kit for Blood).

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

RNA can be extracted within 30 minutes.

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).

RNA_Analysis_SPLIT_mRNA_Extraction_Kit

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.

Fig.2_SPLIT_miRNA_PAGE

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).

SPLIT-Blood_Graphic

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).

 

Workflow

Troubleshooting Guide

 

View the Troubleshooting Guide

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Product Items

Catalog Nr. Product Name
008.48 SPLIT RNA Extraction Kit, 48 extractions
099.48 SPLIT RNA Extraction Kit for Blood, 48 extractions