Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)

3 ARTICLES PUBLISHED IN JoVE

Biology

A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in Plants

Lala Aliyeva-Schnorr¹, Lu Ma^1,2, Andreas Houben¹

¹Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), ²School of Biological and Chemical Sciences, Queen Mary University of London

A protocol is described for the preparation of high-quality mitotic plant chromosome spreads by a fast air-dry dropping method suitable for the FISH detection of single and high copy DNA probes.

Genetics

Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA

Seyyed Hamidreza Hashemipetroudi¹, Ghorbanali Nematzadeh¹, Gholamreza Ahmadian², Ahad Yamchi³, Markus Kuhlmann⁴

¹Genetic and Agricultural Biotechnology Institute of Tabarestan (GABIT), Sari Agricultural Sciences and Natural Resources University (SANRU), ²Department of Industrial and Environmental Biotechnology, National Institute of Genetics Engineering and Biotechnology (NIGEB), ³Department of Plant Breeding and Biotechnology, Gorgan University of Agricultural Sciences & Natural Resources, ⁴RG Abiotic Stress Genomics/ RG Heterosis, Department Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)

Here, we present a protocol for tracing genomic DNA (gDNA) contamination in RNA samples. The presented method utilizes primers specific for the internal transcribed spacer region (ITS) of ribosomal DNA (rDNA) genes. The method is suited for reliable and sensitive detection of DNA contamination in most eukaryotes and prokaryotes.

Biochemistry

Obtaining High-Quality Transcriptome Data from Cereal Seeds by a Modified Method for Gene Expression Profiling

Vito M. Butardo Jr.¹, Christiane Seiler², Nese Sreenivasulu³, Markus Kuhlmann²

¹Department of Chemistry and Biotechnology, Swinburne University of Technology, ²Department of Molecular Genetics, Heterosis Research Group, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), ³Grain Quality and Nutrition Center, Applied Functional Genomics Cluster, International Rice Research Institute

A method for transcriptome profiling of cereals is presented. The microarray-based gene expression profiling starts with the isolation of high-quality total RNA from cereal grains and continues with the generation of cDNA. After cRNA labelling and microarray hybridization, recommendations are given for signal detection and quality control.