Measuring expression of NlKr-h1 by real-time
PCR
N. lugens individuals of every development stage were
collected over a 3-day period for RNA isolation. Samples
consisted of 30-40 of the 1st instar nymphs, 20-30 of the 2nd,
3rd, 4th, and 5th instar nymphs, ten 7-day-old brachypterous
adults, and ten 7-day-old macropterous adults. 20-30
embryos were collected on each day of the 9-day embryonic
development stage. The experiment was repeated 3 times.
Tissues for RT-PCR were dissected from long-wing brown
planthoppers under a Nikon stereo-microscope. Total RNA
was isolated from brown planthoppers at these different
developmental stages or from different tissues using RNAiso
Plus (TaKaRa Bio-Technology Co., Ltd., Dailian, China).