NanoString technology is focused on diagnostic tools for cancer. Because of technological advancements made by NanoString Technologies Inc., it is now possible to perform basic research and translational medicine. The advancements have contributed to significant strides in the medical field.
NanoString Technologies developed the nCounter technology. It is a more sensitive, robust, and easier to use a variety of DNA microarrays. The experts behind https://www.onramp.bio/nanostring explained that the platform can count separate mRNA transcripts. It comes in handy in applications like diagnostics, studying the pathways of gene regulations, and validating the experiments of gene expressions in certain diseases. It can generate translatable data that experts need for clinical use and research.
The nCounter analysis system is the only platform that delivers direct profiling of single molecules in a multiplexed reaction without needing amplification. NanoString technology is considered superior to other RNA analysis techniques for FFPE-derived material. Keep reading to learn more about the use of NanoString technology.
The Benefits of the nCounter Platform
- High flexibility of FFPE and other samples input
- The ability to measure up to 800+ genes in one reaction
- Ability to analyze and detect samples digitally hence reducing the possibility of errors
- No need for polymerase steps or signal amplification
The Benefits of NanoString
- It is amplification-free
- It does not require high-quality RNA. Therefore, it is perfect for scenarios where only low-quality RNA can be accessed. This includes formalin-fixed, paraffin-embedded material
- Low background signal
- It requires small quantities of starting material
- NanoString Technologies provides free data analysis software. The software has user-friendly readouts that are great for non-specialists
- The platform has improved ability to detect low-expression RNA
- Low possibility of introduction bias
The technology uses a different approach from the fluorescence-based expression. It is different from traditional methods. Instead of calculating an intensity score, NanoString technology can profile specific molecules directly. It uses a highly multiplexed reaction and assigns fluorescently-labeled probes to the relevant genes. Probes that are bound to a molecule are counted by a computerized optical lens. Unlike the other methods of gene expression, NanoString does not demand the use of polymerization or pre-amplification. This means that you can avoid amplification bias and the issues it may present. Molecular amplification deviation may cause under or over-amplification of some sequences.
Because of the random nature of its probe-binding, there is a limited likelihood of gene drop-outs. Since it does not depend on fluorophore signal intensity, there are no compensation-based errors.
NanoString minimizes user error by using digital detection and analysis. NanoString assays are extremely robust, reproducible, and sensitive. They are all unique and appropriate for different applications. NanoString technology uses barcodes for the mRNA transcripts of interest. It is possible to assess more than 800 genes per sample and up to 12 samples for every cartridge.
Since NanoString technology doesn’t depend on fluorescence scores or signal intensity, there is little to no bias. The increased accuracy makes it a better option than the other options. The biotin molecules bind mRNA hybridized-reporter strands to the cartridge surface. The barcodes can be detected after being linearized by the induction of electrical gradients.
Individual strands of mRNA can then be identified as representatives of specific genes. All of the captured probes are counted to determine the number of strands that were actually captured in every sample. Because of digital detection and specific direct profiling, it is possible to produce huge data quantities in a short time when necessary.
NanoString technology is used when there is a need for quality control. The data obtained from nCounter Gene Expression experiments is of credible quality. It can safely be used in future statistical analysis.
Unlike the traditional technologies which are based on hybridization, RNAseq does not demand specific probes. It is, therefore, an amazing tool for detecting gene fusions, novel transcripts, and an array of changes that can be difficult to detect.
How to Use the NanoString Platform
The NanoString platform uses two hybridization probes per mRNA molecule. There are probe pairs for every unique gene. Reporter probes hold the signal and the other probes capture the complex. Digital analyzers detect the reporter probes after they are hybridized. There are four unique fluorophores.
The probes are then mixed with RNA. The digital analyzer can scan, identify, and count molecular barcodes in every sample.
In conclusion, the analysis system for NanoString’s nCounter is the first platform to deliver direct profiling of independent molecules in highly multiplexed reactions without amplification. It is superior to the traditional techniques of RNA analysis.