My student has bundles of bee-collected pollen from a phenology study, and would like to get DNA barcode data from them. We have tried following protocols involving liquid N2, and grinding 0.1g of pollen into "fine powder" with mortar and pestle. It never seems to "powderize" but rather turns into a mushy mess. Yields are very low, between 8 and 12ng/uL. Would appreciate any tips on pollen extraction. Post-(semi-successful) powderization, we use the Qiagen DNEasy plant extraction kit.
Pollen has a VERY high lipid and protein content. Try an old school phenol chloroform extraction after grinding in the extraction buffer.
Thank you Katie. We will look into that. Haven't done one of those in years, mostly work with insect & spider tissue, which is usually pretty easy to extract via kits.
I am also using the DNeasy Plant Kit (Mini) with around 0.02g Pollen sample and got DNA yield around 2 to 10 ng/ul but sometines also up to 20ng/ul. Have you tried Katie's suggestion? Does it work better? :-) Or did you find any other improvements? The only thing i altered is to use the pollen sample directly with the beads and half of the buffer volume in a tissue lyser, without freezing...
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