Bio-Fab offer includes DNA and RNA sequencing.
Nanopore technology offers advantages in all reserch areas.
- Genomics and Transcriptomics
- Microbiome-Metagenomics and entire loci sequencing (e.g.: 16S rRNA entire gene)
- Epigenetics and methylation analysis
- AMPLICON SEQUENCING
Amplification of specific targets with a length between 1000-2000 nt with customized protocols.
- 16S AND SHOTGUN METAGENOMICS
16S rRNA sequencing allows the characterization of the prokaryotic microbiome.
Shotgun metagenomics allows the identification of all the genes of all organisms present in a complex sample.
Long and ultra long reads obtained with ONT facilitate genome assembly by providing longer and unambiguously assembled continuous sequences, resulting in fewer contigs.
WGS advantages using TGS:
- Structural variants, breakpoints and repeats identification
- Complete genomes assemblies with de novo or reference-guided approaches
- Direct genome sequencing eliminating PCR bias
- Entire plasmids and viruses sequencing through single reads, eliminating assembly
cDNA-SEQ and Direct RNA-SEQ
- Directly epigenetic modifications identification
Unlike traditional RNA-Seq techniques, ONT allows characterization of cDNA and/or native RNA without fragmentation or amplification steps, thus simplifying analysis and removing potential sources of error.
cDNA seq and RNA seq advantages through TGS:
- Differential gene expression analyisis. The high yield of long reads allows identification of transcriptional isoforms, providing a true reflection of gene expression.
- Fusion transcripts identification. With ONT, read lenght is equal to fragment length, allowing for long and complete transcripts analysis. This minimizes the impact of multimapping, in which short reads align to multiple positions, and allows the complete characterization of transcriptional isoforms and chimeric transcripts.
- RNA viruses sequencing and viral epidemiology analysis. ONT allows complete sequencing of viral RNA in a single read manner, without needed of assembly.
EPIGENETICS AND METHYLATION ANALYSIS
- Nucleotide modifications detection. Nucleotides modifications such as m6A can modulate the activity and stability of RNA molecules. They have been linked to multiple diseases and antibiotic resistance. Unlike traditional technologies, nanopore technology can sequence native RNA molecules whithout the need of amplification or reverse transcription.
Epigenetics, that is the study of hereditary phenotypic changes that don’t cause the alteration of the nucleotidic sequence, plays a key role in gene expression. Since PCR removes the nucleotides modifications, their detection by traditional sequencing technologies requires the use of special library preparation steps (e.g. bisulfite conversion) that damage nucleic acids, thus generating very short sequencing reads. With Nanopore sequencing, PCR is not necessary, allowing the preservation and the direct sequencing of DNA and RNA modifications, without additional library preparation steps.
Nanopore technology allows to perform Adaptive Sampling by sequencing only a region of interest. When the DNA molecule passes through a nanopore, the sequence is decoded in real time. If the sequence doesn’t match the target, sequencing is stopped and the nanopore is free and ready for the next strand. This allows to analyze only the selected sequences, increasing the coverage of the region of interest.