Whole plasmid sequencing is a technique of long-read sequencing used to read the DNA in a plasmid. It makes it possible to sequence millions of DNA simultaneously using the nanopore technology. Although Sanger sequencing is the most popular method, it uses primers for sequencing portions of the DNA. The pieces are then put together into a long sequence. Sanger sequencing does not sequence the whole plasmid. Therefore, this makes whole plasmid sequencing more effective. This article will discuss the benefits and application areas of whole plasmid sequencing.
The Advantages of Whole Plasmid Sequencing
- No Use of Primer
Primers are used in DNA synthesis. They are short fragments in the nucleic acid. These primers need designing and manufacturing, adding the process to the sequencing procedure. Irrespective of the design tools used, there is no guarantee that the primers will perform effectively in sequencing. The good thing about whole plasmid sequencing is that it does not use primers for long-read sequencing data generation. That removes the tedious process of manufacturing primers, leading to a more streamlined sequencing process.
- Longer Reads
As mentioned, Sanger sequencing uses sections of the plasmid instead of the entire loop. On the other hand, whole plasmid sequencing (WPS) uses the entire plasmid for long-read data. In the past, long reads were inaccurate compared to short reads. That means the process of obtaining data was lengthy and costly. However, improvements have introduced whole plasmid sequencing, making ling read faster and more accurate.
- Faster Sequencing
Compared to the method of using primer, whole plasmid sequencing is faster. An entire plasmid is sequenced, helping to obtain data within a day. However, the turnaround time may vary based on the laboratory.
The Application Areas of Whole Plasmid Sequencing
Whole plasmid sequencing is applicable in various fields, including;
- Microbes Identification
Whole plasmid sequencing technique is used in the identification of microbes. Long sequencing methods were banned for using technology to diagnose infectious diseases. However, this new technology has made it possible to identify pathogens quickly. WPS improves patient health by enabling specialists to use the right drugs correctly. Furthermore, WPS is helping identify antibiotic resistance, bacterial evolution, and other aspects of pathogens.
WPS has also opened room for more accurate and faster diagnostics. It is an effective technique for identifying pathogens and detecting disease resistance. Therefore, it has a promising future in the medical industry, with a high need for short diagnostic time and faster treatment.
- Sequencing Difficult Areas
In some areas, it isn’t easy to sequence the genetic material. For example, repetitive sequences and GC-rich areas are challenging to sequence. This is where whole plasmid sequencing works well.
- Exploring Structural Instability
Other methods of plasmid sequencing focus on the treated DNA area and not the rest of the plasmid. However, WPS considers the genetic material’s degradation and the structural changes that happen during duplication. Remember, DNA stability is critical when using plasmids to develop a treatment. Therefore, with WPS, researchers can identify and overcome the instabilities, leading to the development of an effective and safe treatment.
In a Nutshell
Whole plasmid sequencing is invaluable in gene therapy, disease modeling, and drug discovery. It is an accurate, faster, and affordable technique that offers a clearer picture of the plasmid genetic material.