a

b

c

d

e

f

Identification and characterization of genes by RNA-seq

RNA-seq is an NGS-based technique to determine the presence and quantity of messenger RNA in a given sample at a given time, therefore it is used to observe the continuous changes of the cellular transcriptome.

We guide you from the design of the project to the analysis of the results that you can access from our cloud or the information will be delivered to you. We also put our bioinformatics services at your disposal.

RNA-seq Applications:

a. Discovery of new genes for characters of agricultural importance.
b. Quantification of the differential expression of genes.
c. Identification of variants.
d. Functional characterization of molecular markers.
e. Early selection of individuals with interesting agronomic traits in plant variety genetic improvement programs.
f. Genotype-phenotype association analysis.

SNPs identification

A SNP, or single nucleotide polymorphism, is the substitution of one nucleotide for another at a certain position in the genome, as long as this substitution is found in at least 1% of the population.

SNPs are the most widely used markers today because they offer a greater number of advantages than other markers, such as their large number and wide distribution in the genome of all individuals that offers the possibility of building high-density maps required to isolate and studying genes of traits of interest and the possibility of using various automated genotyping methods.

bioSEQs offers different services for detection and validation of SNPs depending on the characteristics of the species in question and the objectives of your project, such as the development of SNPs from:

  • EST sequences
  • Complete resequencing of genomes
  • RNA sequencing
  • Using ddRAD-seq or genotyping by sequencing (GBS)

SNPs Applications:

a. Genotyping of elite varieties of plants and animal breeds to preserve the ownership of the breeder's right.
b. Find SNPs associated with characters of importance.
c. Pedigree Inference and Evaluation.
d. Estimation of genetic values.
e. QTLs mapping.

a

b

c

d

e

a

b

c

d

Identification and typing by whole genome sequencing

When it comes to identification, reference is made to the classification of the organism in question within an already established group or taxon, for its part, the typification refers to a more specific classification, identifying variants that even allow knowing the subspecies. Genome sequencing allows deeper typing, being able to offer information such as antibiotic resistance genes, virulence genes, etc.

This is necessary in the following cases:

1- Identification of strains with little description, low frequency of isolation, or phenotypically atypical.

2- Identification of microorganisms that are not cultivable or difficult to culture.

3- Identification of microorganisms whose biochemical characteristics are not adapted to those of any recognized genus or species, which can sometimes go as far as the description of new pathogens.

4- Taxonomic or phylogenetic studies for epidemiological purposes.

Identification and typing applications by whole genome sequencing:

a. Fungi, bacteria, algae.
b. Biofertilizer registration.
c. Ecological analysis.
d. Difficult to classify microorganisms.

Species identification by DNA metabarcoding

DNA barcoding is a species identification method that uses a short segment of DNA from a specific gene or genes (ITS, 16S or 18S). It is based on the premise that one of these individual sequences can be used as the barcode of the species (ITS Sequence of a fungus) and compared to a library of those sequences, to identify the species in the same way as scanners of supermarkets identify products by their barcode. Metabarcoding is a large-scale taxonomic identification of complex environmental samples that is based on high-throughput sequencing technologies such as Illumina and Oxford Nanopore Technologies, available from our company.

This technology enables researchers to explore microbiome and environmental samples at a much deeper level than was previously possible.

a. Indicators or pests.
b. Verification of raw materials and food ingredients.
c. Study of contaminating microorganisms in food.
d. Stock homogeneity.
e. Biodiversity study.
f. Characterization of the soil microbiome.
g. Monitoring of microbial communities in bireactors.

a

b

c

d

e

f

g

a

b

Detection, tracking and monitoring of microorganisms by qPCR

qPCR, or quantitative PCR, is the most widely used and reliable technique to detect and determine the abundance of microorganisms such as viruses, bacteria, archaea, fungi, and protists in plant or animal tissue samples, soil samples, and other environmental samples.

It is highly specific as it is based on the detection of a nucleic acid segment of the species under study, and it is also a rapid and cost-effective assay.

To quantify the amount of microorganisms present in a given sample, calibration curves are generated from known concentrations of plasmid or genomic DNA. By running our samples and comparing them with this curve, the number of nucleic acid molecules present in the samples of interest can be quantified.

The process consists of the following main stages:

  1. Taking the sample
  2. Isolation of RNA or DNA by a suitable protocol and synthesis of cDNA for RNA samples
  3. Amplification by qPCR with specific primers for the species in question
  4. Analysis of the results and estimation of the abundance of the target DNA in relation to the standard curve
  5. Preparation and delivery to the client of the complete report with the raw data, the characteristics of the test, and the processed results of the analysis.
a. Grapevine
b. Host-pathogen interaction

Characterization of clones or varietal identification by genotyping of microsatellite markers

Microsatellites, or Simple Sequence Repeats (SSRs), are motifs of 1-6 adjacent repeating nucleotides that are found distributed in the genome in both coding and non-coding regions.

The microsatellite technique consists of the PCR amplification of these hypervariable regions using primers for specific regions surrounding the microsatellites.

Its advantages include being relatively abundant, the high level of allelic variation, presenting codominant inheritance, analytical simplicity, and the repeatability of results between laboratories.

With the advent of sequencing techniques, the range of species for which there are primers available to perform this molecular marker technique has expanded.

Its simplicity and high reproducibility make it especially useful for the characterization of clones and varietal identification.

bioSEQs offers a varietal identification service with microsatellite markers for various plant species.

The process consists of the following stages:

  1. DNA isolation of the genotypes under study
  2. Development of the PCR technique with specific primers and separation of the products
  3. Analysis of the results
  4. Delivery of report to the client