16S rRNA of lactic acid bacteria: human breast milk
After PCR amplification, the purified amplicons were subjected to capillary sequencing using the ABI BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, USA). The cycling parameters include pre-holding at 4 C, 1 min at 96 C, 25 cycles at 95 C for 10 s, 50 C for 5 s 62 C for 4 min, and holding at 4 C. The reaction products were subjected to ethanol precipitation before capillary electrophoresis on the ABI 3730xl DNA Analyzer with a 50 cm 96 capillary array, POP7 Polymer, and 3730 Data Collection Software v3.1. The results were finally subjected to base calling and sate quality assessment using the Sequencing Analysis Software v5.4.