Microfluidic Analysis of DNA and RNA

Isolation and purification of DNA/RNA from patients and cells is a standard protocol. Nowadays, microfluidic devices conjugate DNA sequencing and qRT-PCR assisted by syringe pump systems; it is essential to accomplish the purification of DNA or RNA before the sequencing procedure.  Often, these devices perform sample preparation in series with the diagnostic tests. The preparation steps are vital to purify the desired genetic material from potential inhibitors that can interfere with the outcome of the trial. There are various techniques used to selectively capture the nucleic acids at the same time washing out the sources of contamination as proteins, enzymes, lipids, and organic molecules. Liquid and solid support phase extract the target DNA in the above procedures (Fan, 2015)⁠.

 

Syringe pumps provide advantages for DNA/RNA purification

The first DNA microfluidic purification system was a valveless system with micro-sample processing, it was designed for forensic analysis, and it provided several advantages over bench-top instrumentation. The extraction was processed with silica as a solid phase and a syringe pump the flow was driven inside the solid phase, and then the PCR amplification was accomplished. The main advantage is the low-volume during the procedure (Bienvenue et al., 2010)⁠. The closed system evinced the advantages of purification and DNA purification for the amplification of target DNA in an integrated all in one setup. The authors highlighted the potentials of the system and eventually derived in other systems.

 

Why are microfluidics essential for purification of new devices?

DNA and RNA techniques have primarily been used in the molecular diagnosis for research and development; especially the nucleotides purification is required from complex samples like blood. The miniaturization of the micro platform provides the framework the ability to deal with small sample size. The idea behind this approach is to purify the nucleic acids from the rest of the material which may inhibit an accurate diagnosis. DNA/RNA are purified through silica surfaces and in some cases with precipitation; these methods are substantially improving with the implementation of microfluidics.

 

Purification of DNA/RNA in a microfluidic device

Integrated DNA and RNA extraction and purification on an electronic microfluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections and it is recently reported in the literature. The authors automated the procedure in the following assay steps:  sample re-suspension, pre-treatment, lysis, nucleic acid purification, and concentration; all these steps gather into a microfluidic lab-on-a-chip (LOC) cassette that is operated hands-free by a demonstrator setup, providing fluidic and valve actuation. The system evaluates the presence of Gram-positive and Gram-negative cultures in real samples. DNA and RNA is purified through the microfluidic device, and the results are similar to standard extraction with no automate procedures. The proposed setup will help in the development of fast diagnosis and accurate analysis; specifically, the S. aureus and S. pneumoniae were successfully identified after the purification by qRT-PCR (Liesbet Van Heirstraeten et al., 2014)⁠.

 

How Can Chemyx syringe pumps help with DNA/TNA purification?

The Fusion 200 and 4000 are syringe pumps ready to be used in the purification of DNA/RNA by implementation in microfluidics; they provide accuracy fast flow rates and control at low dosages or small volumes.

Finally, the chemyx syringe pumps in conjugation with the purification method reported by Liesbet Van Heirstraeten DNA/RNA can provide faster periods and high quality of the extraction and therefore better quality of the amplification products. It is expected to get results in small times as 60 minutes with full automatization.

 

Concluding remarks

The implementation of Chemyx syringe pumps has been presented for the purification of DNA or RNA in microfluidics. However, the proposal here described is closely related to other techniques as DNA amplification techniques, RNA sequencing from single cells and Lab-on-a-chip devices. It is clear that the implementation of microfluidics will be in vogue in the years to come. The robust tools that the Chemyx syringe pumps allow the development of microfluidic techniques for biological and medical science.

 

Bibliography

Bienvenue, J.M., Legendre, L.A., Ferrance, J.P., Landers, J.P., 2010. An integrated microfluidic device for DNA purification and PCR amplification of STR fragments. Forensic Sci. Int. Genet. 4, 178–186. https://doi.org/10.1016/j.fsigen.2009.02.010

Fan, A., 2015. HHS Public Access 403–411. https://doi.org/10.1007/978-1-62703-134-9

Liesbet Van Heirstraeten, a Peter Spang, b Carmen Schwind, b Klaus S. Drese,*b Marion Ritzi-Lehnert, b Benjamin Nieto, c Marta Camps, c Bryan Landgraf,†c Francesc Guasch, c Antoni Homs Corbera, d Josep Samitier, d Herman Goossens,  a S.M.-K. and T.R., 2014. Integrated DNA and RNA extraction and purification on an automatedmicrofluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections Liesbet. Lab Chip 14. https://doi.org/10.1039/c3lc51339d

 

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