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Larsson, O.

Publications and source records attributed to Larsson, O..

20 records · Page 2Linked to original sources

Normalization of Mass Spectrometry Data (NOMAD)

MotivationiTRAQ reagent-based mass spectrometry (MS) is a commonly used technology for identification and quantification of proteins in biological samples. Such studies are often performed over multiple MS runs, potentially resulting in introduction of MS run bias that could affect downstream analysis. iTRAQ MS data have therefore commonly been normalized using a reference sample which is included in each MS run. We show, however, that such normalization does not efficiently remove systematic MS run bias. A linear model approach was previously proposed to improve on the reference normalization approach but does not computationally scale to larger data. Here we describe the NOMAD (normalization of mass spectrometry data) R package which implements a computationally efficient ANOVA normalization approach with protein assembly functionality.\n\nResultsNOMAD provides the same advantages as the linear regression solution but is more computationally efficient which allows superior scaling to larger sample sizes. Moreover, NOMAD efficiently removes bias which allows for valid across MS run comparisons.\n\nAvailabilityThe NOMAD Bioconductor package: www.bioconductor.org\n\nContactola.larsson@ki.se; carl.murie@ki.se

bioinformatics↗

Polysome-profiling in small tissue samples

Polysome-profiling is commonly used to study genome wide patterns of translational efficiency, i.e. the translatome. The standard approach for collecting efficiently translated polysome-associated RNA results in laborious extraction of RNA from a large volume spread across multiple fractions. This property makes polysome-profiling inconvenient for larger experimental designs or samples with low RNA amounts such as primary cells or frozen tissues. To address this we optimized a non-linear sucrose gradient which reproducibly enriches for mRNAs associated with >3 ribosomes in only one or two fractions, thereby reducing sample handling 5-10 fold. The technique can be applied to cells and frozen tissue samples from biobanks, and generates RNA with a quality reflecting the starting material. When coupled with smart-seq2, a single-cell RNA sequencing technique, translatomes from small tissue samples can be obtained. Translatomes acquired using optimized non-linear gradients are very similar to those obtained when applying linear gradients. Polysome-profiling using optimized nonlinear gradients in HCT-116 cells with or without p53 identified a translatome associated with p53 status under serum starvation. Thus, here we present a polysome-profiling technique applicable to larger studies, primary cells where RNA amount is low and frozen tissue samples.

molecular biology↗