Construction of subtractive cDNA libraries and RT-PCR. (A) Purification of mRNA from A. indica tissues. (B, C) Primary and secondary PCR for immature fruit-enriched (B) and leaf-enriched (C) subtractive cDNA library. (D) Restriction digestion of plasmids isolated randomly from immature fruit-enriched subtractive cDNA library. (E) RT-PCR of cytochrome P450-like gene (a candidate in our library) by using the cDNA made from the total RNA isolated from different tissues of A. indica by our modified method. (F) Restriction digestion releasing the cytochrome P450-like gene fragment from the TA cloning vector (pGEMT-Easy) confirming its successful cloning.