Protocols

A lot of the work in our lab at present involves microarrays and RT-PCR. It can be hard to extract high quality RNA in bulk from Poplar due to the large phenolic content of leaves. These protocols have been extensively tested on a range of Poplar species.

RNA extraction

Bulk extraction. Reliable , yields of 150 µg total RNA from 1 g poplar leaf material, typical purity of 1.8 (A 260 /A 280 )
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Mini version of the above extraction. Good for RT-PCR (but remember to do a DNase treatment!). Yield ~ 20 µg total RNA from 0.04 g young leaf material, typical purity of 1.8 (A 260 /A 280 )
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RNEasy Mini kit extraction for arabidopsis . Yield ~ 30 µg total RNA from 0.06 g leaf material, typical purity of 1.7 (A 260 /A 280 )
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Microarrays

Arabidopsis

Our current Arabidopsis microarray experiments utilise oligo arrays from David Galbraith at Arizona University, USA. Below are our adapted protocols.

cDNA synthesis and post labelling
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Array preparation and hybridisation
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Poplar