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Different protocols and kits used for tissue RNA isolation and DNA isolation

Nowadays, researchers can access both genomic, as well as transcriptomic data, from a single tissue sample because of the advance of NGS or Next Generation Sequencing. The aptitude to isolate high-quality RNA and DNA from a single biological sample is turning out to be more and more important, particularly when the sample yields little amounts of total nucleic acids.

Extracting DNAs and RNAs from tissue samples is mostly exploited for downstream processes, such as library generation, subcloning, real-time quantitative PCR, and conventional Sanger or next-generation sequencing. An important factor in kit selection is the state or type of tissue you are using.

To attain an effective tissue RNA isolation and DNA isolation, researchers have developed a protocol by making use of Beckman Coulter reagents to separate both DNA, as well as RNA, from the same developed tissue samples. The method exploits a proprietary buffer to bind DNA and RNA selectively and reduces the need to divide a lysate before binding and cleaning the nucleic acid sample.

Simultaneous RNA and DNA extraction from the tissue without dividing lysate is one of the protocols used in tissue DNA isolation and RNA isolation. In this isolation method, RNA and DNA were extracted from frozen liver tissues of a mouse. The yield of RNA and DNA was then measured by RiboGreen RNA Assay and Quant-iT PicoGreen dsDNA Assay. It was found that the yield of 32 µg DNA and 66 µg RNA was obtained from a sample of 10 mg of liver tissue of a mouse. The integrity of DNAs and RNAs was evaluated on an Agilent TapeScreen assay and the resulting RIN scores and DIN scores were found to be intact RNA and DNA with RIN 7.4 and DIN 9.2.

Another protocol that was used for tissue RNA isolation and DNA isolation was the extraction of nucleic acids by making use of commercial kits. Researchers used three types of commercial kits for the process, such as:

  1. The Qiagen AllPrep DNA/RNA Micro kit
  2. The MagMAX-96 Total RNA Isolation kit
  3. The Qiagen AllPrep DNA/RNA Micro kit

Each kit uses different technologies for extracting and isolating DNAs and RNAs from the same tissue samples simultaneously. Moreover, each sample comes with detailed instructions from their manufacturers on the effective use of the kits.

All these three commercial kits caused successful tissue DNA isolation and RNA isolation simultaneously. Extractions by making use of Chaos Buffer, TRIzol, and Qiagen AllPrep RNA/DNA Micro kit produced high-quality RNA/DNA consistently. Although the other two kits produced high-quality nucleic acids, as well, they are inconsistent when compared to that of the Qiagen AllPrep RNA/DNA Micro kit. DNA and RNA were effectively isolated with all protocols with the exclusion of the Invitrogen ChargeSwitch removal of phenol phases and inter-phases.

The ChargeSwitch method for isolating DNAs and RNAs uses a technology that is based on magnetic beads. It is dependent on the pH of the nearby buffer, which was likely interrupted by the organic phases due to preliminary extraction steps. High-quality DNA and RNA, as envisaged on agarose gels, was formed with the Qiagen AllPrep DNA/RNA Micro kit and with the Chaos/spin column protocol.

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