Simplex PCR assay for sex determination in mice

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Simplex PCR assay for sex determination in mice.

BENCHMARKS dsDNA was removed from the super-natant (lane 4), which was discarded. The bead-bound dsDNA was denatured in alkaline solution (0.1 M NaOH and 1 mM EDTA) at room temperature for 15 min, and the ssDNA without biotin groups was released in the alkaline solution (lane 5). After neutralization, the ssDNA solution was combined with five volumes of Buffer PB from the MiniElute™ PCR Purific...

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Novel PCR assay for determining the genetic sex of mice.

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Key words: herpes simplex virus, real-time PCR, multiplex real-time PCR assay

要 旨 単 純 ヘ ル ペ ス ウ イ ル ス1型(HSV-1)お よ び2型(HSV-2)の 遺 伝 子 を 同時 に検 出 す るmultiplex real-time PCR法 を 開発 し た. HSV-1お よびHSV-2のgpD領 域 を標 的 と し,プ ラ イ マ ー お よ びTaqMan蛍 光 プ ロ ー ブ を 独 自 に 設 計 し た.検 出感 度 は15copies/tubeで,HSV-1お よ びHSV-2と も15か ら1.5×106 copies/tubeの 範 囲 で 定 量 が 可 能 で あ っ た.real-time PCR法 はPCR法 よ り も103倍 感 度 が 高 く,他 の ヘ ルペ ス属(CMV, EBV, HHV6, HHV-7お よ びVZV) DNAは 検 出 され なか っ た. 以 上 の結 果 か ら,今 回 開 発...

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ژورنال

عنوان ژورنال: BioTechniques

سال: 2005

ISSN: 0736-6205,1940-9818

DOI: 10.2144/05385bm05