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Biochemistry and Molecular Biology Education, Vol. 29 (1) (2001) pp. 21-23
© 2001 IUBMB. Published by Elsevier Science Ltd. All rights reserved.
PII: S1470-8175(00)00065-5

Laboratory exercises

An approach to the critical assessment of the experimental conditions in practical molecular biology:
isolation of plant DNA

Alessio Scarafoni * alessio.scarafoni@unimi.it and Marcello Duranti

Dipartimento di Scienze Molecolari Agroalimentari, Università degli Studi di Milano, Via Celoria, 2, I-20133 Milano, Italy

Abstract

A plant molecular biology laboratory class experiment, based on real lab situations, is proposed to illustrate to the students how the assessment of the experimental conditions and the correct application of the technical details are critical also with an established procedure. During plant genomic DNA isolation and purification, a number of undesired effects, including DNA shearing and formation of secondary compounds, may take place. To simulate such events, we have set up a simple practical laboratory experiment in which the students are asked to isolate plant DNA under three different working conditions, from an optimal to a coarse one. The subsequent analyses on the extracted DNA enable the students to evaluate the influence of these different manipulations on DNA yield and quality. The experiment described can help the students to adopt a critical approach when interpreting the results of common preparative molecular techniques.

Keywords: CTAB; DNA damaging; Genomic DNA; Handling conditions; Quality of isolated DNA

*Corresponding author. Tel.: +39-02-2663662; fax: +39-02-70633062

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