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文献和实验nM 5'biotin end-labeled siRNA under gentle agitation, i.e. a rocking plate (Figure 1a). We utilized the EF1 alpha specific siRNA, EF52, shown previously to direct TGS in human cells (Morris et al. 2004). The EF52 siRNA was constructed with a 5' biotin
Isolating primary B cells from human blood
1. start with freshly drawn human blood not older than 8 hours, treated with an anti-coagulant (use 500ul of 2.7% EDTA for 10ml blood if you are not using EDTA coated tubes when collecting blood) 2. dilute cells with 4 volumes
In situ hybridisation to alpha satellite sequences (chromosome specific)
Alpha satellite sequences, whilst highly repetitive, are specific to each individual chromosome. These sequences flank the centromeres and can present a target measured in megabases. In this protocol a biotin or digoxigenin labelled DNA
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