Construction of 16S rDNA library
The intestinal total bacteria DNA of M. separata
was amplified and purified according
to the above DNA extraction and amplification
process. PCR products were cloned into
pUCm-T according to the manufacturer’s
instructions (Sangon, www.sangon.com) and
then transported into competent cells
(SK2301, Sangon). Plasmid insertions were
screened following the manufacturer’s instructions.
Segment size was checked by PstI
single enzyme cut and agarose gel electrophoresis
to confirm whether the target
fragment was present or not. The cloning
vector was sequenced by the universal primer
m13.