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olfactory epithelia were dissected from three adult female C57BL/6 mice, including tissues attached to the skull and septum. RNA was isolated using the Qiagen RNeasy midi kit (Qiagen, Valencia, CA, USA), including a DNase treatment step. First-strand cDNA was produced from 2.5 μg of RNA in a volume of 50 μl using random hexamers and Invitrogen's Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's recommendations. One-twenty-fifth of the resulting cDNA was used as template in subsequent PCR reactions. PCR amplification biased towards class I olfactory receptors was performed using degenerate primers P26 [17] and classI_R1 (5'-GGRTTIADIRYIGGNGG-3') with an annealing temperature of 44°C. The product was cloned (TA cloning kit, Invitrogen), and individual clones were sequenced. Specific PCR primers used to confirm expression of individual olfactory receptor genes are given in Additional data file 1. Each PCR product was sequenced to confirm that the expected gene and no others had been amplified. Control reactions on a template made by omitting reverse transcriptase gave no product, indicating that the RNA preparation was uncontaminated by genomic DNA."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T12211","span":{"begin":4,"end":13},"obj":"GO:0007608"},{"id":"T12212","span":{"begin":4,"end":23},"obj":"UBERON:0001997"},{"id":"T12213","span":{"begin":50,"end":55},"obj":"UBERON:0007023"},{"id":"T12214","span":{"begin":56,"end":62},"obj":"PATO_UBERON_EXT:female_or_bearer_of_femaleness"},{"id":"T12215","span":{"begin":71,"end":75},"obj":"NCBITaxon:10088"},{"id":"T12216","span":{"begin":87,"end":94},"obj":"UBERON:0000479"},{"id":"T12217","span":{"begin":111,"end":116},"obj":"UBERON:0003129"},{"id":"T12218","span":{"begin":121,"end":127},"obj":"UBERON:0003037"},{"id":"T12219","span":{"begin":129,"end":132},"obj":"CHEBI_SO_EXT:RNA"},{"id":"T12220","span":{"begin":220,"end":225},"obj":"GO_EXT:0004536"},{"id":"T12221","span":{"begin":255,"end":259},"obj":"SO_EXT:cDNA"},{"id":"T12222","span":{"begin":288,"end":291},"obj":"CHEBI_SO_EXT:RNA"},{"id":"T12223","span":{"begin":326,"end":334},"obj":"CHEBI_SO_EXT:hexanucleotide"},{"id":"T12224","span":{"begin":367,"end":388},"obj":"GO_EXT:0003964"},{"id":"T12225","span":{"begin":505,"end":509},"obj":"SO_EXT:cDNA"},{"id":"T12226","span":{"begin":601,"end":610},"obj":"GO:0007608"},{"id":"T12227","span":{"begin":601,"end":620},"obj":"GO_EXT:0004984"},{"id":"T12228","span":{"begin":652,"end":659},"obj":"SO_EXT:0000112"},{"id":"T12229","span":{"begin":717,"end":726},"obj":"GO:0097617"},{"id":"T12230","span":{"begin":764,"end":770},"obj":"SO_EXT:sequence_cloning_process"},{"id":"T12231","span":{"begin":775,"end":782},"obj":"SO_EXT:sequence_cloning_process"},{"id":"T12232","span":{"begin":816,"end":822},"obj":"SO_EXT:sequence_cloned_entity"},{"id":"T12233","span":{"begin":852,"end":859},"obj":"SO_EXT:0000112"},{"id":"T12234","span":{"begin":876,"end":886},"obj":"GO:0010467"},{"id":"T12235","span":{"begin":901,"end":910},"obj":"GO:0007608"},{"id":"T12236","span":{"begin":901,"end":919},"obj":"GO_EXT:0004984"},{"id":"T12237","span":{"begin":920,"end":925},"obj":"SO_EXT:0000704"},{"id":"T12238","span":{"begin":968,"end":979},"obj":"SO_EXT:0000006"},{"id":"T12239","span":{"begin":1023,"end":1027},"obj":"SO_EXT:0000704"},{"id":"T12240","span":{"begin":1111,"end":1132},"obj":"GO_EXT:0003964"},{"id":"T12241","span":{"begin":1170,"end":1173},"obj":"CHEBI_SO_EXT:RNA"},{"id":"T12242","span":{"begin":1208,"end":1219},"obj":"SO_EXT:genomic_DNA"},{"id":"T12243","span":{"begin":1216,"end":1219},"obj":"CHEBI_SO_EXT:DNA"}],"text":"The olfactory epithelia were dissected from three adult female C57BL/6 mice, including tissues attached to the skull and septum. RNA was isolated using the Qiagen RNeasy midi kit (Qiagen, Valencia, CA, USA), including a DNase treatment step. First-strand cDNA was produced from 2.5 μg of RNA in a volume of 50 μl using random hexamers and Invitrogen's Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's recommendations. One-twenty-fifth of the resulting cDNA was used as template in subsequent PCR reactions. PCR amplification biased towards class I olfactory receptors was performed using degenerate primers P26 [17] and classI_R1 (5'-GGRTTIADIRYIGGNGG-3') with an annealing temperature of 44°C. The product was cloned (TA cloning kit, Invitrogen), and individual clones were sequenced. Specific PCR primers used to confirm expression of individual olfactory receptor genes are given in Additional data file 1. Each PCR product was sequenced to confirm that the expected gene and no others had been amplified. Control reactions on a template made by omitting reverse transcriptase gave no product, indicating that the RNA preparation was uncontaminated by genomic DNA."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T12175","span":{"begin":4,"end":13},"obj":"GO:0007608"},{"id":"T12176","span":{"begin":4,"end":23},"obj":"UBERON:0001997"},{"id":"T12177","span":{"begin":50,"end":55},"obj":"UBERON:0007023"},{"id":"T12178","span":{"begin":71,"end":75},"obj":"NCBITaxon:10088"},{"id":"T12179","span":{"begin":87,"end":94},"obj":"UBERON:0000479"},{"id":"T12180","span":{"begin":111,"end":116},"obj":"UBERON:0003129"},{"id":"T12181","span":{"begin":121,"end":127},"obj":"UBERON:0003037"},{"id":"T12182","span":{"begin":601,"end":610},"obj":"GO:0007608"},{"id":"T12183","span":{"begin":652,"end":659},"obj":"SO:0000112"},{"id":"T12184","span":{"begin":717,"end":726},"obj":"GO:0097617"},{"id":"T12185","span":{"begin":852,"end":859},"obj":"SO:0000112"},{"id":"T12186","span":{"begin":876,"end":886},"obj":"GO:0010467"},{"id":"T12187","span":{"begin":901,"end":910},"obj":"GO:0007608"},{"id":"T12188","span":{"begin":920,"end":925},"obj":"SO:0000704"},{"id":"T12189","span":{"begin":968,"end":979},"obj":"SO:0000006"},{"id":"T12190","span":{"begin":1023,"end":1027},"obj":"SO:0000704"},{"id":"T12191","span":{"begin":1208,"end":1215},"obj":"SO:0001026"}],"text":"The olfactory epithelia were dissected from three adult female C57BL/6 mice, including tissues attached to the skull and septum. RNA was isolated using the Qiagen RNeasy midi kit (Qiagen, Valencia, CA, USA), including a DNase treatment step. First-strand cDNA was produced from 2.5 μg of RNA in a volume of 50 μl using random hexamers and Invitrogen's Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's recommendations. One-twenty-fifth of the resulting cDNA was used as template in subsequent PCR reactions. PCR amplification biased towards class I olfactory receptors was performed using degenerate primers P26 [17] and classI_R1 (5'-GGRTTIADIRYIGGNGG-3') with an annealing temperature of 44°C. The product was cloned (TA cloning kit, Invitrogen), and individual clones were sequenced. Specific PCR primers used to confirm expression of individual olfactory receptor genes are given in Additional data file 1. Each PCR product was sequenced to confirm that the expected gene and no others had been amplified. Control reactions on a template made by omitting reverse transcriptase gave no product, indicating that the RNA preparation was uncontaminated by genomic DNA."}

{"project":"2_test","denotations":[{"id":"14611657-10089886-44684104","span":{"begin":664,"end":666},"obj":"10089886"}],"text":"The olfactory epithelia were dissected from three adult female C57BL/6 mice, including tissues attached to the skull and septum. RNA was isolated using the Qiagen RNeasy midi kit (Qiagen, Valencia, CA, USA), including a DNase treatment step. First-strand cDNA was produced from 2.5 μg of RNA in a volume of 50 μl using random hexamers and Invitrogen's Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's recommendations. One-twenty-fifth of the resulting cDNA was used as template in subsequent PCR reactions. PCR amplification biased towards class I olfactory receptors was performed using degenerate primers P26 [17] and classI_R1 (5'-GGRTTIADIRYIGGNGG-3') with an annealing temperature of 44°C. The product was cloned (TA cloning kit, Invitrogen), and individual clones were sequenced. Specific PCR primers used to confirm expression of individual olfactory receptor genes are given in Additional data file 1. Each PCR product was sequenced to confirm that the expected gene and no others had been amplified. Control reactions on a template made by omitting reverse transcriptase gave no product, indicating that the RNA preparation was uncontaminated by genomic DNA."}