Different methods for DNA extraction associated with different storage periods of total saliva in order to obtain genomic DNA for genetic sequencing

• Grant number: 11/22434-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2012
• End date: December 31, 2013
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Lucimara Teixeira das Neves
• Grantee: Thais Francini Garbieri
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

It is well recognized that proper DNA processing is mandatory in order to achieve effective genome sequencing. The first step to this outcome is to choose the starting material from which genetic material will be collected, for example, from which type of cell the DNA will be extracted. In this study, saliva will be used to obtain oral mucosal epithelial cells because the collection is painless, non-invasive, and without any risk of disease transmission. Likewise, saliva does not require specialized personnel to collect and this procedure is well accepted by the majority of the volunteers. Moreover, unlike blood, saliva can be frozen prior to DNA extraction, while blood requires DNA extraction immediately after being collected. Freezing of saliva will be considered in this study because it is well known that freezing saliva for a short period of time may bring benefits, as the freeze can probably inactivate enzymes that are able to degrade DNA. On the other hand, after a certain time of freezing, the genetic material is degraded. This study intends to test the effectiveness of a commercial kit that specifies the preservation of saliva before DNA extraction and that has a preservative substance that is being released in the collection tube of stored saliva. Thus, the aim of this research project is to evaluate the quality and quantity of genomic DNA extracted from the whole saliva with the purpose of direct sequencing, while the specific objectives are to determine whether different storage periods can influence the quantity and quality of DNA with the purpose of direct sequencing to investigate mutations. Also, it is aimed to check whether different DNA extraction protocols can influence these same factors, taking into account the different periods of storage by freezing saliva. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

THAIS FRANCINI GARBIERI; DANIEL THOMAS BROZOSKI; THIAGO JOSÉ DIONÍSIO; CARLOS FERREIRA SANTOS; LUCIMARA TEIXEIRA DAS NEVES. Human DNA extraction from whole saliva that was fresh or stored for 3, 6 or 12 months using five different protocols. Journal of Applied Oral Science, v. 25, n. 2, p. 147-158, 2017-04-00. (09/53848-1, 11/22434-7)