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文献和实验Clonality - X Chromosome Inactivation Assay
can be utilized. 1. Materials DNA sample (see Processing of Microdissected Tissue - DNA-based Analysis) Proteinase K (Sigma) Proteinase K buffer (0.05 M Tris-HCL, 0.001 M EDTA, 1% Tween 20, 0.1 mg/ml proteinase K, pH 8.0) Phenol:chloroform:isoamyl alcohol
Preparation of Microinjection Buffer
EDTA, pH 8.0 (autoclaved) 100 mM NaCl 1 ml of 5 M NaCl (autoclaved) 1x Polyamines 50 microliters 1000x Polyamines mix Autoclaved H2O up to 50 ml NOTE: Prepare fresh and discard unused microinjection buffer.
DIRECT ELISA USING FLUORESCENT SUBSTRATE PROTOCOL
: TBS 250 μl/well; 3x 30 seconds.8. Amplify signal by adding 100 μl/well AttoPhos Fluorescent substrate system, for 5-10 min at RT. 36 mg ofAttoPhos substrate should be mixed with 60 ml of AttoPhos buffer 24 hours prior to use. Make sure well isclean
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