A method for obtaining DNA from compost.
An effective cell lysis method for extraction of bacterial genomic DNA from compost was developed in this study. Enzymatic disruption method, physical-chemical combination method, and commercial kit method were used to extract DNA from compost samples and were compared by analyzing DNA yield and efficient cell lysis. The results showed that all the three methods can be used to extract high-quality DNA from compost, but the enzymatic method had better cell lysis efficiency and DNA yields than others without the use of special equipment and expensive spending. Comparison of different methods for lysing gram-positive bacteria Bacillus subtilis indicated that the enzymatic cell lysis is superior for destroying the gram-positive cell wall. Spin-bind DNA column was used for DNA purification, and the purity of the purified sample was checked by polymerase chain reaction to amplify a region of the 16S rRNA. Results indicated that the part of 16S rRNA were amplified from all the purified DNA samples, and all the amplification products could be digested by the restriction enzyme HhaI.
Wu
Liang
L
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
Li
Fenge
F
Deng
Changyan
C
Xu
Dequan
D
Jiang
Siwen
S
Xiong
Yuanzhu
Y
D001412:Bacillus subtilis / Q000235:genetics; D004269:DNA, Bacterial / Q000302:isolation & purification* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000378:metabolism; D015183:Restriction Mapping; D012987:Soil / Q000032:analysis*
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012987:Soil