Molecular systematics laboratory

The Molecular Systematics Laboratory (LSM) of the Royal Botanic Garden of Madrid (CSIC), was set up in 1997 in order to incorporate new molecular techniques into the RJB's research into plants and fungi biodiversity from three main disciplines:
 
1. Systematic (Taxonomy and Phylogeny)
2. Evolutionary Biology
3. Ecology.
 
Molecular biology applied in systematic studies has contributed to broadening the understanding of evolutionary history (Biogeography, Phylogeography, Evolutionary Ecology, Conservation Biology), and has allowed applied studies such as the identification of emerging pathogenic fungi.
 
Since its creation, LSM has been a key fundamental support in scientific production, which includes a large number of scientific articles in high impact journals.

 

Users

New LSM users will contact Lab Manager that will facilitate access to the facilities, as well as the lab rules.

 
Currently the LSM has users from research groups of the center itself, both researchers, postdoctoral, predoctoral and students. The LSM also welcomes external users from various national and international R&D centers. The interest in using our infrastructure is also notable through programs such as SYNTHESYS (Synthesis of Systematic Resources) integrated infrastructure initiative funded by the European Union. To date, more than 70 external users from other European countries, within the SYNTHESYS program, have developed their research activity at the LSM.
 
 

Techniques

Molecular biology techniques routinely applied in the laboratory:
 
• Extraction of DNA from plants, fungi and bacteria ,

  • Nuclear and cytoplasmic DNA amplification by PCR,
  • Purification of amplified products,
  • Gene cloning,
  • Culture of fungi and bacteria,
  • Fingerprinting techniques (RFLP, AFLP, microsatellites, RADs),
  • Fragmentation of nucleic acids as a previous step for libraries generation,
  • Quality control of NGS libraries.

 

 

Equipment


The LSM has the following equipment:


 
 
• Covaris M220 ultrasonic. Focused ultrasonication equipment allows the fragmentation of DNA or RNA samples for analysis by NGS techniques. The process is carried out at 500 kHz (ultrasonic range) outside the audible range, avoiding inconvenience for operators, without heat-induced bias, with high sample recovery and avoiding cross contamination. It is a device that allows the fragmentation process of the genetic material of the samples to be carried out with high precision and reproducibility, and is used in all cases in which the enzymatic fragmentation of nucleic acids is not viable.

• Bioanalyzer 2100 Agilent. Automated processing system, for multiple samples simultaneously (12-96), for the separation of DNA and RNA by high-resolution digitized capillary electrophoresis, from a very small amount of sample (1-2 microliters), which offers an interpretation in peaks by an electropherogram in addition to the traditional interpretation of gels. It allows an effective control of the integrity of the DNA obtained after the extraction processes, as well as an effective quality control of the DNA libraries, presenting among other advantages its speed, low toxicity and high sensitivity, reliability and reproducibility.

• Mastercycler X50S thermal cyclers. With silver block, for 96-well plates that allow optimization of PCR amplification protocols through the configuration of 2D gradients, being able to adjust specifically and for each position, both the denaturation temperature, and the hybridization temperature during it cycle.

• NanoDrop® 1000 is a micro-drop spectrophotometer that measures nucleic acid concentrations from 1 µl of sample, with great accuracy and reproducibility.
Qubit® Fluorimeter for highly specific quantification of nucleic acids.
• TissueLyser ® II. Sample homogenization equipment, for the rapid and efficient rupture of tissues prior to the extraction and purification of nucleic acids. The rupture of tissue and / or cells is achieved through the use of steel or glass balls inside the tube sample.

• Virtis Benchtop K 6KBTEL-85 freeze dryer. Sublimation drying equipment used as a preliminary step to the extraction and purification of DNA from biological samples.




Fee

The billing of the services provided will be made to the Research Group Managers or Principal Investigators Responsible for Projects according to the current rate by type of user.

 





Real Jardín Botánico
Real Jardín Botánico, CSIC. Plaza de Murillo, 2. Madrid E-28014 (ESPAÑA). Tel. +34 91 4203017. FAX: +34 91 4200157
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