title: cDNA library sequencing in the frontal gland of soldiers in Nasutitermes takasagoensis by 454 GS Junior pyrosequencing
organism: Nasutitermes takasagoensis
attributes.tag: sample_name
attributes.value: DRS001664
attributes.tag: sample comment
attributes.value: Nasutitermes takasagoensis collected from Ishigaki Island were used. For construction of a primary cDNA library, the frontal glands were dissected from the heads of soldiers. The RNA may have been from frontal gland cells and around tissues such as muscle and fat bodies. First strand cDNA was synthesized using an oligo(dT)18 anchor primer that included an XhoI restriction site and an adaptor sequence with an EcoRI restriction site according to the Gubler–Hoffman method. The cDNA was ligated into a pBluescript II SK(+) vector digested with EcoRI and XhoI restriction enzymes. The vector was transfected into E. coli DH10B competent cells. The plasmid was purified by phenol–chloroform extraction and ethanol precipitation and then diluted in TE buffer. The primary cDNA library was amplified because the quantity was insufficient for cDNA library preparation for GS Junior sequencing. The primary cDNA library was transformed into E. coli HST08 Premium Competent Cells. The amplified plasmid was used as a sample DNA to prepare a shotgun library for GS Junior Titanium Series.