Young and healthy maize leaves were collected from the two parental lines and 261 RILs at the seedling stage, frozen in liquid nitrogen, and stored at –80 °C. Total genomic DNA was extracted using the cetyltrimethylammonium ammonium bromide (CTAB) method [48 (link)]. All samples with a suitable concentration and quality were used for library construction.
The library was constructed using the SLAF sequencing method described previously [49 (link)]. Two restriction enzymes, Hpy166II and HaeIII (New England Biolabs, Ipswich, Massachusetts, USA), were selected to digest the genomic DNA into 414- to 464-bp fragments. The ends of completely digested fragments were repaired into blunt-ended DNA and the 5' ends were phosphorylated. An adenine base was added to the 3' end of the fragment, after which the duplex index sequencing adapter was connected to the A-tailed fragment. The target fragment was amplified, purified, and sequenced on an Illumina HiSeq 2500 platform (Illumina Inc., San Diego, California, USA).
The library was constructed using the SLAF sequencing method described previously [49 (link)]. Two restriction enzymes, Hpy166II and HaeIII (New England Biolabs, Ipswich, Massachusetts, USA), were selected to digest the genomic DNA into 414- to 464-bp fragments. The ends of completely digested fragments were repaired into blunt-ended DNA and the 5' ends were phosphorylated. An adenine base was added to the 3' end of the fragment, after which the duplex index sequencing adapter was connected to the A-tailed fragment. The target fragment was amplified, purified, and sequenced on an Illumina HiSeq 2500 platform (Illumina Inc., San Diego, California, USA).
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