DNA and RNA Sizing and Quantification

QC and DNA sizing

Separation and sizing of nucleic acids is critical for many applications including:

  • NGS library assessment.
  • Size distribution assessment of high molecular weight DNA.
  • Large DNA fragment analysis.
  • Assessment and quantification of total RNA, messenger RNA, cDNA and microRNA.
  • Reverse genetics analysis, mutation detection.
  • SSR, genotyping and PCR fragment analysis.
  • CRISPR/Cas9 gene editing.

Finding the right system that can cover a broad range of applications and perform size separation easily and accurately on large numbers of samples is not easy.

With the unique ability to house two different gel matrices, the Fragment Analyzer can run not only more samples, but also different types of samples unattended. The operational software automates the gel switching and filling between different applications, eliminating unnecessary wait times. More than 20 Gel Kits are available for a wide range of applications.

The Fragment Analyzer accelerates nucleic acid analysis workflow through the automation of key steps such as gel loading and sample injection. It not only speeds up lab processes, the instrument also enables more consistent and reproducible results than competitive platforms. By covering the widest fragment range - from microRNA to high molecular weight genomic DNA - the Fragment Analyzer does more and offers unique features not found in other instruments.

Nucleic Acid Quantification

Many techniques of cellular and molecular biology require the ability to quantify dsDNA and RNA in large numbers of samples at sensitivities that only require a small amount of the total sample. Isolation of plasmids from bacterial cultures, genomic DNA from mammalian cells, cDNA synthesis for library production, and quantitation of PCR products for diagnostic purposes all require the direct quantitation of nucleic acids. Also, many biochemical studies that involve the growth kinetics of cell cultures or cell cycle studies require normalisation by DNA content.

The most commonly used method for the determination of nucleic acid concentration is the determination of absorbance at 260 nm). This method, while quite adequate for many situations, can suffer from the interfering absorbance of contaminating molecules. Many of these contaminates which include nucleotides, RNA, EDTA and phenol are commonly found in nucleic acid preparations. As a result, several fluorescent staining techniques have been developed to alleviate many of the problems associated with absorbance at 260 nm.

Biotek plate readers are a perfect platform for a wide range of direct detection UV absorbance quantification protocols and fluorescent detection protocols in medium to high throughput plate-based format down to as little as 1-2 uL.