Otimização e validação de um produto biotecnológico utilizado na extração de DNA.

Abstract

Deoxyribonucleic acid, known as DNA and an organic compound whose molecules contain as genetic instructions that coordinate the development and functioning of all beings, and transmit as hereditary characteristics of each living being. Its main function is stored as necessary information for a construction of proteins from RNAs. In biotechnology as information present in the molecules are analyzed and from the analysis and possible diagnostic modifications in the cell, it is necessary to extract the genetic material from the cell. Initially a product was developed in a house that can extract DNA from blood samples and biopsy in natura. In a new phase of development phase, start optimizing this, the idea is to improve the product and validate the product. The reagents already developed followed protocols adapted from Tost et al. (2003) and Miller et al. (2000), generating solutions such as: cell lysis and nuclear lysis (safety solutions for cell and nuclear membrane respectively), purification solution and rehydration solution (solution to resuspend the genetic material), as they are primordial solutions for extraction. In this work, more solutions were added to the kit to extract the genetic material from different samples, for which several protocols were studied, for the types of samples are very different and optimization of a protocol specific for each sample to one of the most importantly presented in this work, unqualified, protocols and solutions to extract DNA from in natura tissue, blood, paraffin biopsy, bacteria and protozoa. The extracted genetic material was evaluated by spectrophotometry and visualization in agarose gel, and the material was still tested in a PCR. A creation of a new image for the product was also developed by obtaining a didactic product following its purpose and at a lower cost compared to other commercial kits. The product was internally validated by UNISC research groups. The objective of this project was to improve the "CPTBio DNA Extraction" kit by expanding its use for paraffin-labeled biopsies, gram negative bacteria and protozoa, thereby achieving a DNA extraction product developed at home at a lower cost, generating DNA of quality and amount compatible for use in various molecular techniques.