Method for recovery of intact DNA for community analysis of marine intertidal microbial biofilms.
A protocol is described for rapid DNA isolation from marine biofilm microorganisms embedded in large amounts of exopolysaccharides. The method is a modification of the hot phenol protocol used for plants tissues, where nonexpensive and easily available enzymes were used. The method is based on the incubation of biofilm biomass samples in an extraction buffer mixed with phenol preheated at 65 degrees C. The procedure can be completed in 2 h and up to 20 samples can be processed simultaneously with ease and DNA of excellent quality, as shown by successfully amplification of polymerase chain reaction (PCR) products. DNA was recovered from a range of intertidal marine biofilms with varying amounts of exopolysaccharides.
Narváez-Zapata
José A
JA
Departamento de Microbiología Ambiental y Biotecnología, Programa de Corrosión del Golfo de México, Universidad Autónoma de Campeche, Av. Agustín Melgar, Col. Lindavista, C.P.24030, Campeche, Campeche, Mexico. janavae@mail.uacam.mx
Rodríguez-Avila
Norma
N
Ortega-Morales
Benjamín O
BO
D001419:Bacteria / Q000145:classification / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods; D018441:Biofilms / Q000145:classification; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D009792:Oceans and Seas; D016133:Polymerase Chain Reaction / Q000379:methods; D014871:Water Microbiology
D004269:DNA, Bacterial