chloroplast genome of Martinella obovata: leaf sample
Five micrograms (5 g) of genomic DNA were fragmented using a Covaris S-series sonicator to obtain DNA fragments of approximately 300 bp. The sonicator parameters for 50 L of sample were: 10% duty factor, 200 cycles per burst, 50 s treatment time and intensity five. Short-insert libraries were constructed using the NEBNext DNA Library Prep Master Mix Set and NEBNext multiplex oligos for Illumina (New England Biolabs Inc., Ipswich, USA). We verified expected size selection by agarose gel electrophoresis of library products against a size standard Promega 100 bp ladder (Promega Corporation, Madison, USA). We determined DNA library concentration on an Applied Biosystems 7500 Real-Time PCR System using the Kapa Library Quantification Kit (Kapa Biosystems Inc., Wilmington, USA) and/or using fluorometry on Qubit 2.0 (Life Technologies). The DNA library was pooled together with other 15 samples sequenced on an Illumina HiSeq 2000 system (paired-end) (Illumina Inc., San Diego, USA).