端分子标记 16 Cd冰12001)
1≈
原位杂交流程 探针的标记 ■取材 ■切片:石蜡切片冰冻切片 杂交前预处理 杂交后洗去未结合的及非特异性结合的探 针 ■探针的检测
原位杂交流程 ◼ 探针的标记 ◼ 取材 ◼ 切片:石蜡切片 冰冻切片 ◼ 杂交前预处理 ◼ 杂交 ◼ 杂交后洗去未结合的及非特异性结合的探 针 ◼ 探针的检测
探针的标记 Hybridization robes DNA RNA Oligonucleotide Transcription from Ori Cell-based DNA cloning or PCR insert DNA cloned in suitable vectors Chemical synthesis Characteristics Normally double-stranded Single-stranded Single-stranded of starting 0. 1 kb to hundreds of kb for usually up to a usually material conventional DNA clones, 0.1 few thousand 15-50nt kb to >20 kb for PCR products nucleotides long long Run-off transcription Labeling Usually by DNA polymerase. End labeling(e. g based DNA strand synthesis from cloned DNA by polynucleotide (see Figure 5.2) (see Figure 5.4) kinase; see Figure 5.3)
探针的标记