characteristics.text: SAMN00117445
characteristics.characteristic: External Id
characteristics.text: Michigan State University
characteristics.characteristic: INSDC center name
characteristics.text: 2010-11-08T09:37:16.060Z
characteristics.characteristic: INSDC first public
characteristics.text: 2013-06-12T09:08:11.970Z
characteristics.characteristic: INSDC last update
characteristics.text: SRS118978
characteristics.characteristic: INSDC secondary accession
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Generic
characteristics.characteristic: NCBI submission model
characteristics.text: Generic.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: SRS118978
characteristics.characteristic: SRA accession
characteristics.text: Seeds of Momordica charantia collected at 18 days after pollination. Independent male flowers were used for hand pollination of female flowers. Embryos were dissected from seeds of fruits collected at specific DAP and frozen immediately in liquid nitrogen. Embryos were stored at -80°C until use in RNA isolation. Total RNA was extracted from developing bitter melon seeds was ground to a fine powder in liquid nitrogen using Trizol reagent (Invitrogen) according too the manufacturer’s protocol.mRNA were purified from 1 mg of total RNA by two passes through oligo-dT cellulose columns by use of the Illustra mRNA purification kit (GE Healthcare). cDNA libraries were constructed using SMART PCR cDNA synthesis kit (Clontech). First-strand cDNA was synthesized with 150 ng mRNA in a volume of 10 ml using the provided SMART II primer, a modified CDS III/ 3’ cDNA synthesis primer (5’-AAGCAGTGGTATCAACGCAGAGTGGCCGAGGCGGCCGACATGTTTTGTTTTTTTTTCTTTTTTTTTTVN-3’) and Superscript II reverse transcriptase (Invitrogen). Double stranded cDNA was prepared by PCR (18 cycles) using 2 ml of the first-strand reaction in a 50 ml reaction volume. Following Proteinase K treatment, four PCR reactions were pooled before SfiI digestion and size fractionated on the provided CHROMA SPIN-400 column. Only fractions containing fragments larger than 500 bp were collected, precipitated, and resuspended in TE buffer.
characteristics.characteristic: description
characteristics.text: Momordica charantia
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3673
characteristics.characteristic: organism
characteristics.text: Non-normalized cDNA from Momordica charantia seeds
characteristics.characteristic: title