LIBEST_028195
External Id
SAMN02146484
External Id
Stazione Zoologica Anton Dohrn
INSDC center name
2013-06-22T23:05:03.773Z
INSDC first public
2015-05-20T15:03:17.827Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
The SSH experiment was carried out for two reciprocal FORWARD and REVERSE subtractions. Shallow library (FORWARD subtraction, S) was carried out with shallow growing plant mRNA as a tester pool and deep growing plant mRNA as a driver pool. At the same time, in the Deep library (REVERSE subtraction, D)deep mRNA was used as a tester pool and shallow mRNA as a driver pool. The construction of the forward and reverse SSH libraries was performed using the PCR-select cDNA subtraction kit (Clontech, Palo Alto, CA) and following the procedure suggested by the manufacturer. Two micrograms of polyA+ mRNA from each of the tester and driver populations were converted to double-stranded cDNA by reverse transcription followed by digestion with RsaI to produce shorter blunt ended fragments. The digested tester cDNA was subdivided into two populations, each of which was ligated with a different adaptor (ADAPTOR 1 5'-CTAATACGACTCACTATAGGGCTCGAGCAGCGGCCGCCCGGGCAGGT-3'; ADAPTOR 2R 5'-CTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAGGT-3') provided in the cDNA subtraction kit (Cat: No:637401).Clones were sequenced with the Sanger method using a ABI 3730xl automated DNA sequencers (Applied Biosystems, USA).
description
Posidonia oceanica
http://purl.obolibrary.org/obo/NCBITaxon_55489
organism
leaf
http://purl.obolibrary.org/obo/PO_0025034
tissue-type
LIB EST_Pooc SSH
title
pCR 2.1-TOPO
vector