characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-02-20T17:01:10Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T17:08:38Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2335566
characteristics.characteristic: External Id
characteristics.text: ETH_Zurich
characteristics.characteristic: INSDC center name
characteristics.text: 2014-02-20T17:01:10Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T17:08:38Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS393760
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2226:pool of nine mice
characteristics.characteristic: Submitter Id
characteristics.text: 8 week
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000034
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Immunized with NP-CGG (chicken gamma globulin (CGG) conjugated to 4-hydroxy-3-nitrophenylacetyl)
characteristics.characteristic: clinical information
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: All animal experiments were performed under the guidelines and protocols approved by the Basel-Stadt cantonal veterinary office (Basel-Stadt Kantonales Veterin_ramt Tierversuchsbewilligung #2582). Female BALB/c mice (n=9, Charles Rivers Laboratories) 6 to 8 weeks old were housed under specific pathogen-free conditions and were maintained on a normal chow diet. Nine BALB/c mice (8-10 weeks old, Charles River, specific-pathogen free) were immunized with 50 ug alum-precipitated chicken gamma globulin (CGG) conjugated to 4-hydroxy-3-nitrophenylacetyl (NP, NP-CGG, BioCat). Mice were sacrificed 14 days post-immunization (dpi) and their spleens and bone marrow (from femurs and tibia) were harvested. Antibody-secreting CD138-positive cells were enriched from spleen and bone marrow 14 dpi as previously described (Reddy et al., 2007, Nat Biotech) Total RNA extraction from the pooled ASC population was performed using the TRIzol Plus RNA Purification Kit (Life Technologies), according to the manufacturer's protocol and concentration was determined on a Nanodrop 2000c Spectrophotometer. Enriched total RNA was homogeneously mixed, aliquoted and stored at -80 degrees C Direct Addition method: The Illumina TruSeq Universal Adapter constituted the 5' portion of the forward primers, while the IgG-reverse primer contained the Illumina index primer (Index 4 [Replicate DA1] and 5[Replicate DA2]), thereby directly adding Illumina adapter sequences to PCR products (Fig. 1). 10 PCR reactions were run in parallel with the standard PCR conditions mentioned above, which is equivalent to 500 ng total RNA. Thusly prepared libraries were concentrated and purified as stated above.
characteristics.characteristic: description
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: spleen, bone marrow
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: BALB/c
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000602
characteristics.characteristic: strain
characteristics.text: pool of nine mice
characteristics.characteristic: title