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DNA 300 bp to 700 bp was isolated using a MinElute Gel Extraction Kit (Qiagen).

The full length BbKst gene (7375 bp) was isolated using RACE and genome walking.

Samples were then run out on a 1.5% agarose (Sigma), 0.5X TBE gel, and DNA 300 bp to 800 bp was isolated using a MinElute Gel Extraction Kit (Qiagen).

Pooled DNA sample was run out on a 1% agarose (Sigma, USA), 0.5 × TBE gel, and DNA fragments 200 bp to 500 bp was isolated using a Min Elute Gel Extraction Kit (Qiagen, Germany).

DNA fragments in the range of 300 500 bp were isolated using the MinElute Gel Extraction Kit (Qiagen).

Regulatory sequences upstream of the initiation codon of PpNAC2 (of 661 bp) and PpNAC3 (1115 bp) were isolated using a Genome Walking approach.

After the digestion (enzyme MseIwas used), polymerase chain reactions (PCR) and purification of genomic DNA, fragments(with indices and adaptors) of 330 ~ 380 bp were isolated using Gel Extraction Kit (Qiagen) and subjected to PCR amplification (barcode2 was added).

Samples were then separated by electrophoresis through a 1.5% agarose, 0.5× TBE gel, and DNA fragments from 300 to 700 bp were isolated using a MinElute Gel Extraction Kit (Qiagen).

Using degenerate primers designed on the basis of conserved domain of insect deduced SID-1 amino acid sequences, a partial sequence (231 bp) was obtained from S. exigua by RT-PCR, and a longer 939 bp sequence was isolated using 3'-RACE with the specific primers on the 231 bp PCR fragment.

DNA was isolated using standard extraction protocols.

Total RNA was isolated using Isogen (Wako).