Mosquito Blood Meal Analysis Protocol

Mosquito abdomens were removed for blood meal analysis using disposable razor blades. DNA was extracted from the abdomen of blood-fed mosquitoes using DNAzol BD (Molecular Research Center, Cincinnati, OH) according to the manufacturer’s recommendation with modifications as described elsewhere [21 (link), 32 (link), 33 (link)]. Isolated DNA from the mosquito blood meals was used as a DNA template in subsequent polymerase chain reaction (PCR) assays with primers based on mitochondrial cytochrome b sequences of avian and mammalian species. The thermal cycling conditions were as described earlier [21 (link), 32 (link), 33 (link)]. The Veriti Dx Thermal Cycler, or GeneAmp PCR System 9700 (Applied Biosystems, Foster City, CA) were used to perform PCR assays, and PCR products were sequenced directly in cycle sequencing reactions using the sequencer 3730xl DNA Analyzer (Applied Biosystems) at the Keck Sequencing Facility (Yale University, New Haven, CT). Sequence annotation and analyses were conducted using ChromasPro version 1.7.5 (Technelysium Pty Ltd., Tewantin, Australia), and compared to the sequences available at the GenBank DNA sequence database of the National Center for Biotechnology Information using the BLAST search (BLASTN) [34 ].

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Molaei G., Armstrong P.M., Graham A.C., Kramer L.D, & Andreadis T.G. (2015). Insights into the recent emergence and expansion of eastern equine encephalitis virus in a new focus in the Northern New England USA. Parasites & Vectors, 8, 516.

Publication 2015

Veriti Dx Thermal Cycler GeneAmp PCR System 9700 sequencer 3730xl DNA Analyzer ChromasPro version 1.7.5

Abdomen Assays Avian Blood Blood analysis Cytochrome b Mammalian Mitochondrial Mosquito Polymerase chain reaction Primers

Corresponding Organization :

Other organizations : Connecticut Agricultural Experiment Station, New York State Department of Health

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Variable analysis

independent variables

Mosquito abdomens were removed for blood meal analysis using disposable razor blades.
DNA was extracted from the abdomen of blood-fed mosquitoes using DNAzol BD (Molecular Research Center, Cincinnati, OH) according to the manufacturer's recommendation with modifications as described elsewhere [21, 32, 33].

dependent variables

Isolated DNA from the mosquito blood meals was used as a DNA template in subsequent polymerase chain reaction (PCR) assays with primers based on mitochondrial cytochrome b sequences of avian and mammalian species.
PCR products were sequenced directly in cycle sequencing reactions using the sequencer 3730xl DNA Analyzer (Applied Biosystems) at the Keck Sequencing Facility (Yale University, New Haven, CT).
Sequence annotation and analyses were conducted using ChromasPro version 1.7.5 (Technelysium Pty Ltd., Tewantin, Australia), and compared to the sequences available at the GenBank DNA sequence database of the National Center for Biotechnology Information using the BLAST search (BLASTN).

control variables

The thermal cycling conditions were as described earlier [21, 32, 33].
The Veriti Dx Thermal Cycler, or GeneAmp PCR System 9700 (Applied Biosystems, Foster City, CA) were used to perform PCR assays.

Annotations

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