647荧光原位杂交(FISH)成像试剂盒

647荧光原位杂交(FISH)成像试剂盒，ReadiLink™ iFluor® 647 FISH Fluorescence Imaging Kit，是使用647标记的 FISH 探针通过原位杂交标记靶 DNA 的便捷工具。更多视频请关注视频号【艾维缔】。哔哩哔哩【IVDSHOW】。抖音【军哥聊表观】。

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染料探针

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17316

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荧光原位杂交（FISH）技术是检测生物标本中特定核酸靶点的有效工具。通过荧光染料标记的核酸探针与标本的互补序列杂交，可实现核酸靶标的原位检测。647荧光原位杂交(FISH)成像试剂盒是使用 iFluor® 647 标记的 FISH 探针通过原位杂交标记靶 DNA 的便捷工具。该试剂盒提供 Taq DNA 聚合酶，通过聚合酶链反应 (PCR) 将 iFluor® 647-dUTPs 结合到目标 DNA 中。我们专有的 iFluor® 染料比传统的荧光标签更亮、更耐光照，可提供所需的分辨率和信号。

图 1. 使用 iFluor® 647-dUTP 标记的端粒探针在分裂期 HeLa 细胞中进行端粒定量荧光原位杂交。探针使用647荧光原位杂交(FISH)成像试剂盒制作。

操作步骤

使用前，将所有组分解冻至室温，涡旋充分混合。

注：以下方案可作为标准DNA FISH的通用指南。请根据自己的实验进行方案的修改或优化。

1.配制如下反应混合液，见表1

每个反应每孔的试剂组成

成分	体积 (25 µL/反应)	最终浓度
FISH反应混合物 (2X)	12.5 µL	1X
上游引物, 10 µM	0.25-2.5 µL	0.1-1.0 µM
下游引物, 10 µM	0.25-2.5 µL	0.1-1.0 µM
模板DNA	1-5 µL	优化浓度
iFluor 647-dUTP	2.5 µL
dNTP 混合物	1 µL
水，无核酸酶	25 µL

2.通过缓速涡旋，然后用短暂的离心机小心地混合试剂

3.在qPCR仪中置板，按下表2操作

表2 .热循环参数

参数	聚合酶活化	PCR (30-40 个周期)
	持续温度	变性	退火	延长
温度	95 °C	95 °C	55-65 °C	68-72 °C
时间 (m:ss)	0:20	0:30	1:00	1:00

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647荧光原位杂交(FISH)成像试剂盒
17316
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