Genotyping Arabidopsis T-DNA lines v1

This protocol is used for genotyping Arabidopsis seedlings to test for the presence of a transfer DNA (T-DNA) insertion. By using two primer sets it is possible to determine whether a seedling is homozygous, heterozygous or azygous for an insertion in the predicted genomic location. To identify lines with T-DNA insertions in a gene of interest, you need the Arabidopsis Genome Identifier (AGI) number corresponding to the genomic locus (e.g. RCS1A = AT1G67090), then visit the Salk Institute T-DNA Express site to find all the mapped insertions at your locus of interest. Genotyping primers have been pre-designed for each T-DNA line, these can be retrieved from the Salk Institute T-DNA primer site, and ordered at any supplier of DNA oligonucleotides before starting the protocol. In the US T-DNA lines can be purchased from the Arabidopsis Biological Resource Center (ABRC) and in the UK and EU from the European Arabidopsis Stock Center (NASC). Recommended reading http://signal.salk.edu/tdnaprimers.2.html Setting up the PCR reaction Genotyping is performed with the Phire Direct PCR Mix, this includes the polymerase, nucleotides and salts necessary for amplification. We use the “dilution protocol” which involves taking a small leaf disk and homogenizing it in dilution buffer using a gel tip (see manufacturer’s instructions for more details.)

Genotyping Arabidopsis T-DNA lines v3

This protocol is used for genotyping Arabidopsis seedlings to test for the presence of a transfer DNA (T-DNA) insertion. By using two primer sets it is possible to determine whether a seedling is homozygous, heterozygous or azygous for an insertion in the predicted genomic location. To identify lines with T-DNA insertions in a gene of interest, you need the Arabidopsis Genome Identifier (AGI) number corresponding to the genomic locus (e.g. RCS1A = AT1G67090), then visit the Salk Institute T-DNA Express site to find all the mapped insertions at your locus of interest. Genotyping primers have been pre-designed for each T-DNA line, these can be retrieved from the Salk Institute T-DNA primer site, and ordered at any supplier of DNA oligonucleotides before starting the protocol. In the US T-DNA lines can be purchased from the Arabidopsis Biological Resource Center (ABRC) and in the UK and EU from the European Arabidopsis Stock Center (NASC). Recommended reading http://signal.salk.edu/tdnaprimers.2.html Setting up the PCR reaction Genotyping is performed with the Phire Plant Direct PCR Mix, this includes the polymerase, nucleotides and salts necessary for amplification. We use the “dilution protocol” which involves taking a small leaf disk and homogenizing it in dilution buffer using a gel tip (see manufacturer’s instructions for more details.)

Genotyping Arabidopsis T-DNA lines v1

This protocol is used for genotyping Arabidopsis seedlings to test for the presence of a transfer DNA (T-DNA) insertion. By using two primer sets it is possible to determine whether a seedling is homozygous, heterozygous or azygous for an insertion in the predicted genomic location. To identify lines with T-DNA insertions in a gene of interest, you need the Arabidopsis Genome Identifier (AGI) number corresponding to the genomic locus (e.g. RCS1A = AT1G67090), then visit the Salk Institute T-DNA Express site to find all the mapped insertions at your locus of interest. Genotyping primers have been pre-designed for each T-DNA line, these can be retrieved from the Salk Institute T-DNA primer site, and ordered at any supplier of DNA oligonucleotides before starting the protocol. In the US T-DNA lines can be purchased from the Arabidopsis Biological Resource Center (ABRC) and in the UK and EU from the European Arabidopsis Stock Center (NASC). Recommended reading http://signal.salk.edu/tdnaprimers.2.html Setting up the PCR reaction Genotyping is performed with the Phire Direct PCR Mix, this includes the polymerase, nucleotides and salts necessary for amplification. We use the “dilution protocol” which involves taking a small leaf disk and homogenizing it in dilution buffer using a gel tip (see manufacturer’s instructions for more details.)

Paralelos diagramáticos na obra de Louis I. Kahn: do edifício Richards Medical Research Laboratories ao Salk Institute for Biological Studies

Richards Medical Research Laboratories e Salk Institute for Biological Studies: dois complexos laboratoriais projetados pelo profícuo arquiteto americano Louis I. Kahn e, construídos, respectivamente, nos intervalos temporais de 1957 a 1964 e de 1959 a 1965. Não obstante, a programática geral que rege e dá origem aos edifícios é, de certa maneira, o único ponto em comum no que tange a contextualização teórico-prática e físico-histórica das obras. A partir da inquietação apresentada e mediante a constatação das diferenças que alçaram estes dois projetos a grandes exemplares da arquitetura projetada e construída nos Estados Unidos na segunda metade do século XX, surge a proposição de traçar paralelos que tornassem possíveis comparações tanto teóricas e aplicadas, quanto gráficas e diagramáticas. Dessa maneira, o presente trabalho apoiou-se em duas linhas de desenvolvimento complementares que constituem, primeiramente, de levantamentos histórico-práticos da arquitetura de Louis I. Kahn, bem como de seus contemporâneos e, em última instância, na transformação das informações coletadas em diagramas arquitetônicos que conferem a essas bases novos significados e morfologias. Em suma, os resultados obtidos buscaram transformar relações condensadas e estáticas sobre os dois edifícios contrapostos – Richards Medical Research Laboratories e Salk Institute for Biological Studies – em produtos gráficos que trazem maior dinamismo às intrincadas variáveis que compõe as obras, sejam elas: contextuais, morfológicas, conceptivas, estruturais, infaestruturais e teóricas