Pilot progress in DNA isolation and amplification from the material stored at the LWS herbarium

Abstract

The isolation of DNA from the herbarium specimens deposited at the LWS herbarium (State Museum of Natural History of the NAS of Ukraine, Lviv, Ukraine) has been tested using the column-based protocol. The isolated DNA has been amplified using different nuclear and plastid primers. The yield of obtained total DNA showed no significant dependence from the year of collection and plant family of studied specimens. In general, the obtained DNA of LWS specimens had medium yield (mean – 56.47 ng/µL) but relatively low purity (mean 260/230 value – 0.85 units and mean 260/280 value – 1.66 units). The success of DNA amplification for old herbarium material varied from 12.5 % to 91.1 % depending on applied primers. The trnL P6 Loop primers demonstrated the best performance (91.1 % successful amplification), but due to short resulted DNA fragments, it was not possible to purify the product for further processing. UniPlant primers performed the worst, and only 12.5 % of samples taken from the LWS herbarium (excluding controls) were successfully amplified. In general, nuclear primers, except for UniPlant, demonstrated a better success rate (mean – 31.5 %) during the work with samples taken from the LWS herbarium. Meanwhile, the plastid primers, except for trnL P6 Loop, showed slightly lower amplification success (mean – 26.8 %).

Keywords

• herbarium specimens
• plant DNA barcoding
• DNA extraction methods
• degraded DNA
• LWS herbarium