18S RNA gene sequences were downloaded from the National
Center for Biotechnology Information (NCBI) (Table 1). From the
resulting alignment with Clustal W (Thompson, Gibson, Plewniak,
Jeanmougin, & Higgins, 1997) integrated into BioEdit 7.0 (Hall,
1999), a specific primers/probe set was designed by using Primer
Express software (Applied Biosystems): FISH ALLERGEN F (50-GGC TCA TTA AAT CAG TTA TG-30), FISH ALLERGEN R (50-CCG AGT TAT CTA GAG TCA-30) and FISH ALLERGEN PROBE (50-6-FAM-CCG TAC
TTG GAT AAC TGT GGC AAT TC-BHQ1-30). The PCR reactions were carried out in a total volume of 20 lL containing 100 ng of DNA template, 10 lL of TaqMan Fast Advanced Master Mix (Applied Biosystems), primers and probe and molecular
biology grade water (Eppendorf) to adjust to the final volume.
18S RNA gene sequences were downloaded from the National
Center for Biotechnology Information (NCBI) (Table 1). From the
resulting alignment with Clustal W (Thompson, Gibson, Plewniak,
Jeanmougin, & Higgins, 1997) integrated into BioEdit 7.0 (Hall,
1999), a specific primers/probe set was designed by using Primer
Express software (Applied Biosystems): FISH ALLERGEN F (50-GGC TCA TTA AAT CAG TTA TG-30), FISH ALLERGEN R (50-CCG AGT TAT CTA GAG TCA-30) and FISH ALLERGEN PROBE (50-6-FAM-CCG TAC
TTG GAT AAC TGT GGC AAT TC-BHQ1-30). The PCR reactions were carried out in a total volume of 20 lL containing 100 ng of DNA template, 10 lL of TaqMan Fast Advanced Master Mix (Applied Biosystems), primers and probe and molecular
biology grade water (Eppendorf) to adjust to the final volume.
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