title: Differential male salivary gland genes-A3
status: Public on Apr 08 2004
contact.city: Stillwater
contact.first: Richard
contact.middle: C.
contact.last: Essenberg
contact.email: richard.essenberg@okstate.edu
contact.state: OK
contact.address: 246 NRC
contact.department: Department of Biochemistry and Molecular Biology
contact.country: USA
contact.institute: Oklahoma State University
contact.phone: 405 744-6189
organism: Dermacentor andersoni
description: Differentially expressed bands from the differential display gels were cut out, eluted and amplified.  Successfully amplified ones were cloned into pDrive (Qiagen).  The cDNA fragment clones were denatured in 0.5N NaOH and 20mM EDTA at 65oC for 1 hour, then spotted onto nylon membranes by a Seiko model X 5000 cartesian robot (Seiko Instruments Inc., Japan) to create the macroarray.  Each cDNA fragment was applied to three corners of a 4.5 mm square on the 16 x 24 pattern.  The fourth corner of the square was left blank for a control. The positive controls used were tick genomic DNA from D. andersoni and two cDNA fragments (standards 1 and 2) that appeared to be expressed in all three groups of experimental ticks from the differential display The negative controls were buffer and pBluescript vector. These clones were amplified with the A1 + A3 primer pair.