title: Ct 5k cDNA array  v1.0
status: Public on Dec 31 2013
contact.city: Guangzhou
contact.first: SH
contact.last: Shi
contact.email: lssssh@mail.sysu.edu.cn
contact.address: 135 Xin Gang Xi
contact.country: China
contact.institute: Sun Yat-Sen University
organism: Ceriops tagal
manufacture_protocol: Mono-clones were collected from a root cDNA library and a leaf SSH library, and the cDNA insertion was amplified by hot-start PCR using the following program: hot-start at 94℃ and a 4 minute hold for pre-denaturation of the template; 32 cycles of 1 minute at 94℃, 30 seconds at 56℃ and 1.5 minutes at 72℃. The M13F/M13R primer pair (5’- GTA AAA CGA CGG CCA GT -3’/ 5’- AAA CAG CTA TGA CCA TGT TCA -3’) was used to prime the reactions. The PCR products were purified using the Multiscreen-PCR 96-well purification system (Millipore, Bedford, MA), dried down, resuspended in 50% DMSO before loaded onto 384-well plates. A total of 3909 cDNA from the root cDNA library and 1330 cDNA from the leaf SSH library were printed onto the poly-L-lysine modified slide (Full Moon Biosystems, Inc.) in duplicate using the GeneMachines Omingrid 100 Arrayer (GeneMachine, San Carlos). Each cDNA chip composes of 48 blocks. Within each block, additional cDNA fragments from six C. tagal housekeeping genes (actin 11, actin 2, ubiquitin 3, ubiquitin 4, 18s rRNA and β-tubulin used as positive controls) and one animal gene (serving as negative control), and the pDNR-LIB vector and 50% DMSO (for background signal monitoring) were also included.