Steps to generate dna barcode of insects
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DNA extraction is the
critical first step in generating DNA barcodes
and can be a rate-limiting step in very large
barcoding studies. Consequently, a DNA
extraction method that is rapid, easy to use,
cost-effective, robust enough to cope with
range of qualities and quantities of tissue,
and can be adapted to robotic systems will
provide the best method for high-throughput
production of DNA barcodes. We tested the
performance of a new commercial kit
(prepGEM), which uses a novel, streamlined
approach to DNA extraction, and we
compared it with two other commercial kits
(ChargeSwitch and Aquapure), which differ in
their method of DNA extraction. We compared
performance of these kits by measuring
percentage of polymerase chain reaction
(PCR) success and mean PCR product yield
across a variety of arthropod taxa,
whichincluded freshly collected, ethanol-
preserved, and dried specimens of different
ages. ChargeSwitch and prepGEM performed
equally well, but they outperformed Aquapure.
prepGEM was much faster, easier to use, and
cheaper than ChargeSwitch, but
ChargeSwitch performed slightly better for
older (> 5-yr-old) dried insect specimens.
Overall, prepGEM may provide a highly
streamlined method of DNA extraction for
fresh, ethanol-preserved, and young, dried
specimens, especially when adapted for high-
throughput, robotic systems.
critical first step in generating DNA barcodes
and can be a rate-limiting step in very large
barcoding studies. Consequently, a DNA
extraction method that is rapid, easy to use,
cost-effective, robust enough to cope with
range of qualities and quantities of tissue,
and can be adapted to robotic systems will
provide the best method for high-throughput
production of DNA barcodes. We tested the
performance of a new commercial kit
(prepGEM), which uses a novel, streamlined
approach to DNA extraction, and we
compared it with two other commercial kits
(ChargeSwitch and Aquapure), which differ in
their method of DNA extraction. We compared
performance of these kits by measuring
percentage of polymerase chain reaction
(PCR) success and mean PCR product yield
across a variety of arthropod taxa,
whichincluded freshly collected, ethanol-
preserved, and dried specimens of different
ages. ChargeSwitch and prepGEM performed
equally well, but they outperformed Aquapure.
prepGEM was much faster, easier to use, and
cheaper than ChargeSwitch, but
ChargeSwitch performed slightly better for
older (> 5-yr-old) dried insect specimens.
Overall, prepGEM may provide a highly
streamlined method of DNA extraction for
fresh, ethanol-preserved, and young, dried
specimens, especially when adapted for high-
throughput, robotic systems.
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