"In-gel patch electrophoresis:" a new method for environmental DNA purification.
Most of the microorganism species are largely untapped and could represent an interesting reservoir of genes useful for biotechnological applications. Unfortunately, a major difficulty associated with the methods used to isolate environmental DNA is related to the contamination of the extracted material with humic substances. These polyphenolic compounds inhibit the DNA processing reactions and severely impede cloning procedures. In this work, we describe a rapid, simple, and efficient method for the purification of genomic DNA from environmental samples: we added a chromatography step directly embedded into an agarose gel electrophoresis. This strategy enabled the DNA extraction from various environmental samples and it appeared that the purity grade was compatible with digestion by restriction enzymes and polymerase chain reaction (PCR) amplifications.
Roh
Changhyun
C
School of Chemical Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea.
Villatte
Francois
F
Kim
Byung-Gee
BG
Schmid
Rolf D
RD
D044002:Bryophyta / Q000235:genetics; D004247:DNA / Q000302:isolation & purification*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D019015:Geologic Sediments / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012722:Sewage / Q000382:microbiology; D012988:Soil Microbiology
D012722:Sewage; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific