LIBEST_022984
External Id
SAMN00165533
External Id
Mount Desert Island Biological Laboratory
INSDC center name
2008-04-14T00:00:00Z
INSDC first public
2024-10-07T21:09:35.507Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
ERS21120385
SRA accession
LEE-1
cell-line
Egg cases of the Little Skate (Leucoraja erinacea) were obtained from wild-caught egg-laying females kept at the Mount Desert Island Biological Laboratory, Salisbury Cove, ME 04672, U.S.A. Embryos were staged according to a scheme developed for Scyliorhinus canicula, the Lesser Spotted Dogfish (J. Exp. Zool. 1993. 267:318-336). The cell line was derived from a single embryo by mincing the head and body. Cells were grown in culture as described by (Hwang, J-H., Parton, A., Czechanski, A., Ballatori, N., and Barnes D.. 2008. Arachidonic Acid-Induced Expression of the Organic Solute and Steroid Transporter-beta in a Cartilaginous Fish Cell Line. Comparative Biochem. and Physiology, in press). Using TRIzol reagent, total RNA was isolated from 0.5 x10e8 cells which had been passaged 10-14 times. The primary cDNA library was constructed from mRNA and directionally cloned into pCMV-SPORT6.1. The host bacterium was DH10B T1 (T1 and T5 phage resistant). cDNA construction, normalization and quality control were performed by Invitrogen. The average insert size was 1.9 kb and the titer was 8.5 x10e6 cfu/ml. Inserts were single-pass sequenced from the 5' end by the University of Washington High-Throughput Genomics Unit, Department of Genome Sciences, Seattle, Washington. Traces were trimmed and prepared at the Mount Desert Island Biological Laboratory, Maine for submission to dbEST (GenBank) using trace2dbest software (Parkinson, Anthony and Baxter, unpublished). This expressed sequence tag project was supported by NIH grant Number 1 RO1 019732 from the National Center for Research Resources (D.W. Barnes, PI), the Woodcock Foundation (D.W. Barnes, PI) and NIH grant P20-RR016463 for Research Resources (P.H. Hand, PI).
description
28
dev-stage
Leucoraja erinaceus
http://purl.obolibrary.org/obo/NCBITaxon_7782
organism
single embryo
tissue-type
Little Skate (Leucoraja erinacea) embryo cell line 1 (LEE-1): 5' sequences of ESTs
title
pCMV-SPORT6.1
vector