Discover SPLIT RNA Extraction Kit

SPLIT RNA Extraction Kit -
Purify RNA to the highest grade!

The SPLIT RNA Extraction Kit enables fast and highly efficient extraction of high-quality, high-purity RNA from various biological samples, including cell culture, animal and plant tissue, and fluid samples. The obtained RNA is ideal for seamless library preparation for Next Generation Sequencing and other demanding applications such as full-length reverse transcription, sample preparation for microarray analysis, or RT-qPCR. SPLIT recovers the complete RNA size range, including small RNAs (<200 nt). Additionally, large and small RNA-enriched fractions can be extracted by following a supplemental protocol.

SPLIT RNA Extraction Kit (Cat. No. 008) offers universal, species-independent RNA extraction from a broad range of materials, including animal and plant tissue, cell cultures, liquid samples, and FFPE samples. SPLIT’s streamlined protocol delivers high-purity RNA within 30 minutes using Phase Lock Gel tubes for rapid, convenient, and safe phase separation.

Exceptional Yields and High-quality RNA Extraction

SPLIT RNA Extraction Kit consistently yields high-quality RNA across diverse sample types, resulting in high RNA Integrity Number (RIN) quality scores. Extraction from cell cultures typically achieve a RIN of 10 and a 28S / 18S rRNA ratio of 2.7, indicating highly intact RNA. Extractions from tissue samples usually result in RNA with a RIN of 8.0 – 9.5 (Figure 1). Beyond quality, SPLIT RNA Extraction Kit demonstrates high extraction efficiency, with typical yields of extracted RNA from mouse liver ranging from 4.0 – 4.5 µg total RNA / mg tissue.

Figure 1 | The SPLIT protocol enables the extraction of high-quality RNA. Gel-like representation of Agilent Bioanalyzer traces. RNA from mouse liver stored in RNAlater was extracted either as total RNA (lane 1) or as large and small RNA-enriched fractions (lanes 2 and 3). A control sample was extracted following a TRIzol protocol (lane 4). All samples have a RIN of 8.2 – 8.3 (not applicable for small RNA-enriched fraction).

Flexible RNA Isolation of Total RNA or Size-enriched RNA Fractions

SPLIT RNA Extraction Kit effectively extracts RNA of all size ranges, including small RNAs (≥17 nt). 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-enriched fraction (cutoff at ~150 nt), with the option to obtain the small RNA-enriched fraction separately (Figure 1). Efficient recovery of siRNAs and miRNAs down to 17 nt in total RNA or the small RNA-enriched fraction has been shown in spike-in experiments with small RNA markers (Figure 2).

Figure 2 | Separation of SPLIT RNA samples on a polyacrylamide gel, demonstrating the splitting of large and small RNA-enriched fractions at a threshold of ~150 nt. The total RNA sample comprising small and large RNA is shown as a comparison. The homogenate was spiked with a miRNA marker to assess efficiency of miRNA recovery. Small RNAs down to at least 17 nt are efficiently recovered in the total RNA sample and in the small RNA-enriched fraction.

The SPLIT RNA Extraction Kit contains reagents for the isolation of total RNA or the large RNA-enriched fraction from 48 samples, or small and large RNA-enriched fractions from 24 samples.

Step1:

Total time:
30 min

The sample is homogenized in an isolation buffer that is highly chaotropic to facilitate effortless and complete solubilization.

The sample is transferred to a phase lock gel column which contains a special gel matrix that acts as a barrier between the organic and aqueous phase based on the density differences.

Acidic phenol and acidic buffer are added to create a monophasic solution, which is essential for the efficient separation of genomic DNA into the organic phase. Chloroform is added and phases are cleanly separated.

After centrifugation the gel acts as a seal between the phases.

The aqueous phase containing the RNA can be easily decanted and no carry-over of the organic phase will take place.

Depending on the amount of isopropanol added, either the total RNA (1.75x isopropanol) can be extracted, or the RNA can be split into a large (0.33x isopropanol) and a small RNA-enriched fraction (1x isopropanol to the flow-through of the large RNA-enriched fraction).

The RNA is precipitated onto a silica column. For the total RNA, the entire RNA will precipitate onto the silica carrier. For the large RNA-enriched fraction, RNA with a lower limit of about 150 nt will bind, whereas the small RNA will be in the flow-through.

The flow-through of the large RNA-enriched fraction contains the small RNAs which didn´t bind to the silica column. After the addition of 1x isopropanol, it can be precipitated and purified onto a new silica column.

10 – 50 µl of Elution Buffer or Storage Buffer are added to the silica membrane to elute the RNA bound to the silica carrier.

RNA extraction is finished and the RNA is ready for anydownstream application.

Frequently Asked Questions

Access our frequently asked question (FAQ) resources via the buttons below.

Please also check our General Guidelines and FAQ resources!

How do you like the new online FAQ resource? Please share your feedback with us!

Troubleshooting Guide

View the Troubleshooting Guide

Problem	Likely cause	Comments and suggestions
No or poor phase separation	Cooled phase-lock gel or insufficient centrifugation	Ensure that phase-lock gel tubes are equilibrated at room temperature prior to the extraction Ensure that centrifugation is carried out at +18 °C or at room temperature Increase centrifugation time or repeat centrifugation step
Incorrect cut-off	Volume of isopropanol addition	Measure volume of aqueous phase to add correct amount of isopropanol (0.33x, 1.75x, or 1x vol depending on the desired RNA fraction)
Degraded RNA	RNA source	Avoid freeze-thaw cycles of your sample Freeze starting material quickly in liquid nitrogen or in appropriate buffer
Degraded RNA	RNase contamination	Avoid speaking over opened tubes and wear gloves Follow protocol closely and work quickly
Low absorption ratios (i.e., peak at 230 nm)	Contamination with organic solvents, salts, or metal ions	Ensure appropriate blank solution (e.g., Storage Buffer (SB) contains EDTA which can influence absorption measurements) Ensure to remove all residual GuSCN by following protocol closely (washing steps and dry spin) Optionally increase the number of washing steps
Low or no RNA yield	RNA remains on spin column	Increase incubation time Perform second elution Optionally pre-heat Elution Buffer (EB) to 60 °C prior to elution Ensure Wash Buffer (WB) has been diluted with 100 % ethanol as indicated on bottles
Low or no RNA yield	Spin column was overloaded	Reduce quantity of starting material (a maximum of 10 µg of RNA can be bound to the spin column)
Problem in downstream application	Salt or ethanol carry-over during elution	Ensure Wash Buffer (WB) has been diluted with 100 % ethanol as indicated on bottles Increase number of washing steps Ensure dry spin to remove all traces of ethanol

User Guide – update 07.10.2022

SPLIT Supplementary Protocol: Purification of RNA from FFPE Samples – upload 18.07.2018

Explore our dedicated FFPE RNA extraction kit: SPLIT One-step FFPE RNA Extraction Kit

Safety Data Sheet

Request Safety Data Sheet

If you need more information about our products, please contact us through support@lexogen.com or directly under +43 1 345 1212-41.

Cat. No.	Product Name
008.48	SPLIT RNA Extraction Kit, 48 extractions

First Time User? We’re excited to offer you an exclusive introductory offer.

Need a web quote?

You can generate a web quote by Register or Login to your account. In the account settings please fill in your billing and shipping address. Add products to your cart, view cart and click the “Generate Quote” button. A quote in PDF format will be generated and ready to download. You can use this PDF document to place an order by sending it directly to sales@lexogen.com.

Web quoting is not available for countries served by our distributors. Please contact your local distributor for a quote.

SPLIT RNA Extraction Kit
Add to cart

SPLIT RNA Extraction Kit -
Purify RNA to the highest grade!

Check the FAQs

Get your special offer

SPLIT RNA Extraction Kit

Performance

Workflow