GH_TMO, LIBEST_017827 (cDNA)

Overview
Library NameGH_TMO
Unique NameLIBEST_017827
OrganismGossypium hirsutum (Upland Cotton)
TypecDNA
Properties
Property NameValue
Vector typeplasmid
VectorpCMV-SPORT6.1
Cloning site1: EcoRI; 2: NotI
Tissue typeImmature ovules
Developmental stageImmature ovules (-3 to 3 days post anthesis, DPA) with or without fibers
CommentsG. hirsutum L. Texas Marker-1 (TM-1) plants were grown in a greenhouse at the Texas A&M University and in the field at the USDA-ARS, New Orleans. The bolls were tagged and harvested at 3 days before anthesis (-3DPA), the day of anthesis (0 DPA), and 3 days post anthesis (3DPA) from a pool of ~20 plants. Total RNA was extracted from immature ovules or fiber-bearing ovules. An equal amount of total RNA from three samples was mixed and sent to Invitrogen Corp. (Carsbad, California) for full-length cDNA library construction. Messenger RNA was isolated from the total RNA using a filter syringe containing oligo (dT). The first-strand cDNA was synthesized from 15 ug mRNA using SuperScriptTMIII reverse transcriptase, and the second-strand cDNA was synthesized using E. coli RNase H, DNA polymerase I, and DNA ligase. The double-stranded cDNA was blunt-ended using T4 DNA polymerase, digested with NotI, size-selected using agarose gel electrophoresis, and directionally cloned into the NotI-EcoRV sites of the pCMV-SPORT6.1. The ligated products were transformed using ELECTROMAXTMDH10BT1 cells. The library contained 4.2x106 cfu with ~100% of colony growth rate. It is estimated that over 60% clones contained full-length cDNA inserts with an average insert size of 1.53-kb. A total of 51,072 colonies were arrayed in duplicate sets each with 133 384-well plates for sequencing and storage, respectively.
Cross References
External references for this cDNA
DatabaseAccession
NCBI BioSampleLIBEST_017827