Attached
Infected chronic wounds are polymicrobial in nature which include a diverse group of aerobic and anaerobic microorganisms. Majority of these communal microorganisms are difcult to grow in vitro. DNA fngerprinting methods such
as polymerase chain reaction-denaturation gradient gel electrophoresis (PCR-DGGE) facilitate the microbial profling
of complex ecosystems including infected chronic wounds. Six diferent DNA extraction methods were compared
for profling of the microbial community associated with chronic wound infections using PCR-DGGE. Tissue debris
obtained from chronic wound ulcers of ten patients were used for DNA extraction. Total nucleic acid was extracted
from each specimen using six DNA extraction methods. The yield, purity and quality of DNA was measured and used
for PCR amplifcation targeting V2–V3 region of eubacterial 16S rRNA gene. QIAGEN DNeasy Blood and Tissue Kit
(K method) produced good quality genomic DNA compared to the other fve DNA extraction methods and gave a
broad diversity of bacterial communities in chronic wounds. Among the fve conventional methods, bead beater/
phenol–chloroform based DNA extraction method with STES bufer (BP1 method) gave a yield of DNA with a high
purity and resulted in a higher DGGE band diversity. Although DNA extraction using heat and NaOH had the lowest
purity, DGGE revealed a higher bacterial diversity. The fndings suggest that the quality and the yield of genomic DNA
are infuenced by the DNA extraction protocol, thus a method should be carefully selected in profling a complex
microbial community