LC+YC : Rumen contents LC-YP-4 : fattening male lamb
Total genome DNA was extracted using the CTAB/SDS method. The 16S rRNA genes were amplified using hypervariable V3-V4 region PCR primers (341F: 5-CCTAYGGGRBGCASCAG-3; 806R: 5-GGACTACNNGGGTATCTAAT-3). All PCR reactions were carried out with 15 L of PhusionHigh-Fidelity PCR Master Mix (New England Biolabs, Ipswich, MA, USA), 0.2 M of forward and reverse primers and about 10 ng template DNA. After amplification, PCR products were mixed with the same volume of SYBR green (QIAGEN Inc., Alameda, CA, USA) containing buffer and then operated electrophoresis on 2% agarose gel. Samples with bright main strip between 400450 bp were purified with GeneJET Gel Extraction Kit (Thermo Fisher Scientific (China) Ltd., Shanghai, China). Sequencing libraries were generated using NEB NextUltraDNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA, USA) following the manufacturers recommendations.
SUB8925435
LC-YP-4