Random Lab Methods

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Rapid Isolation of RNA from Conifer Needles

Conifer_RNA_prep. Conifers join a long list of 'recalcitrant' plants that are difficult for RNA extraction, and fail using "traditional" RNA extraction kits. We use a modification of the method by Tai et al, 2004.

Isolation of poly(A) mRNA with Sera-Mag Oligo(dT) beads

mRNA_Prep. There are many methods for isolating mRNA from total RNA. We have had excellent results with Sera-Mag Oligo(dT) beads. We use this approach for constructing Illumina mRNA-Seq libraries, but it should be useful for any application that demands rRNA-depleted mRNA.

Preparation of strand-specific mRNA-Seq libraries with the Illumina TruSeq RNA Sample kit

directional-RNAseq_Prep. There are many methods available for making strand-specific mRNA libraries. We chose to adapt one of the most reliable methods identified by Levin et al. (2010) - dUTP labelling followed by dUTP degredation - for use in the Illumina TruSeq mRNA kit. This method produces mRNA-Seq libraries that are highly enriched for the complementary sequence of the native mRNA.

Rapid Isolation of DNA from Conifer Needles

DNA_Seq Prep. Conifers join a long list of 'recalcitrant' plants that are difficult for DNA extraction. We use a modification of the Fast-Prep method.

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