RNA Isolation in 7 Minutes – Go!

blog / Molecular Biology December 28 2020

Total RNA is the starting point for a plethora of applications that can help to answer questions about the mechanisms of human and plant disease, gene regulation, embryonic development, transcriptional responses to treatments and stimuli, and more. Regardless of the application in hand – whether it be cDNA synthesis for reverse-transcription PCR, RNA-seq, northern blotting or otherwise, sufficient amounts of high-quality RNA will be a requisite.

There are many approaches to total RNA isolation, such as organic extraction protocols based on guanidinium thiocyanate, phenol and chloroform, solid phase extraction with spin columns or magnetic bead-based protocols, and direct lysis methods that deploy buffers to break open samples and stabilise nucleic acids for downstream applications without further purification. Each of these methods has its merits and drawbacks.

Choosing an RNA Isolation Method

A number of considerations may be relevant when choosing an RNA isolation method, including: speed and hands-on time needed, scalability and automation potential, safety i.e., use of toxic substances that require special disposal, equipment needed, purity, concentration and yield of isolated material.

The sample itself, e.g., presence of cell walls (e.g., fungi, plants), the application and its requirements for quantity, concentration and purity, and the availability of lab resources, e.g., equipment, will usually dictate the choice of RNA isolation method.

Wouldn’t it be ideal if there was one approach that ticked all of the boxes?

Simple and Fast RNA Isolation From Any Sample With Direct-Zol

Direct-zol RNA Kits from Zymo Research are RNA purification kits that provide a streamlined workflow for the purification of up to 100 µg (per prep with Direct-zol RNA Miniprep Plus Kit) of high-quality RNA directly from virtually any sample type stored in TRI Reagent®, TRIzol®, or similar.

The Direct-zol workflow (Figure 1) is simple and it can be done in 7 minutes!

Figure 1. Direct-zol Workflow.

Simply apply a prepared* sample in TRI Reagent directly to the Zymo-Spin Column and then bind, wash, and elute the RNA.

*Certain sample types, e.g., plants, fungi and certain other microbial samples, will require an extra unbiased mechanical cell lysis step to remove cells walls or minimise the bias that may occur due to inconsistent sample lysis for microbiome studies.

High Quality, Small (17 nt) and Large RNAs From Single Cells

With Direct-zol kits, total RNA, including small RNAs (17-200 nt), is effectively isolated from virtually any sample types (as shown in Figure 1 above).

In addition, the Zymo-Spin silica-based binding column is uniquely designed to capture unbiased total RNA down to picogram amounts (Direct-zol RNA Microprep Kit), making it a feasible option for single-cell total RNA isolation as shown in Figure 2 below.

Figure 2. Total RNA purified from 104 down to a single human epithelial cell with the Direct-zol RNA Microprep kit and detected by RT-qPCR for the beta-actin gene.

The Direct-zol workflow bypasses the need for manual phase separation, precipitation, and post-purification steps, leaving you with an elution of high-quality RNA that is suitable for sensitive downstream applications including cDNA synthesis for reverse-transcription PCR, transcriptome analysis by RNA-seq, RNA hybridisation and arrays, and more.

Is Direct-Zol the Solution for You?

The Direct-zol comes in a number of formats and sizes to meet the varied needs of our customers. The table below provides an overview of the kits in the range. Since the kits are compatible with virtually any sample type and yield high-purity RNA from as little as a single cell without the need for any lab specialised equipment, they are a feasible option in many projects.

If you would like to discuss how Direct-zol might simplify and streamline your RNA isolation needs then please get in touch with us for an informal chat where we can guide you. You are also welcome to test the kit yourself with a free sample. You can enquire about this by writing to us at info@nordicbiosite.com.

Direct-zol Products Overview

Cat. No. Product Name Binding Capacity Elution Volume Isolation Format No. Cells required Weight Tissue Required
BioSite-R2061 Direct-zol RNA Microprep 10 µg 6 µl Spin-Column ≤ 1×106 ≤ 5 mg
BioSite-R2051 Direct-zol RNA Miniprep 50 µg 25 µl Spin-Column ≤ 5×106 ≤ 25 mg
BioSite-R2071 Direct-zol RNA Miniprep Plus 100 µg 50 µl Spin-Column ≤ 1×107 ≤ 50 mg
BioSite-R2055 Direct-zol-96 10 µg 10 µl 96-Well Plate ≤ 1×106 ≤ 5 mg
BioSite-R2101 Direct-zol-96 MagBead 10 µg 50 µl Magnetic Beads ≤ 1×106 ≤ 5 mg

Related Blog Posts:

The Lowdown on Real-Time PCR – Part 1

The Lowdown on Real-Time PCR – Part 2

Gene Expression Analysis: RNA-Seq or Real-Time PCR?

One-Step RT-PCR

RNA-Seq 1: Introduction