Dot-blot hybridization conditions for the establishment and optimization of.docVIP

 PAGE \* MERGEFORMAT 9 Dot-blot hybridization conditions for the establishment and optimization of [Abstract] purposes dot the establishment and optimization of hybridization conditions. Methods positively charged nylon membrane for dot-blot hybridization of each step of the optimization, including pre-hybridization solution, hybridization solution, washed and closed and the liquid-step reaction such as time, and used to establish the conditions for the final detection of glycosyltransferase gene peptide : N-acetyl-galactosyltransferase 2 (pp-GalNAcT2) in the K562, NB4, NIH3T3, and SGC7901 cell lines in the differential expression of membrane hybridization conditions to verify the feasibility. Resulted in the creation and optimize the conditions for hybridization of the membrane, the end of a clear and reliable. Conclusion The simple dot blot techno-economic and practical. [Keywords:] dot-blot hybridization; Glycosyltransferase; gene The establishment and optimization of the dot blot prerequisite [Abstract] Objective To establish and optimize the dot blot technique.Methods We used the positively charged nylon membranes to optimize experiments for the dot blot including prehybridization solution, hybridization solution, washing buffer, blocking solution et al, and the times of every step. In order to validate its feasibility, we analyzed the expression of ppGalNAc-T2 in different tumor cell lines by dot blot assay using the established hybridization system.Results We established and optimized the dot blot technique which had reliable results and low background.Conclusion The established dot blot technique is practicable and economical. [Keywords:] dot blot; glycosyltransferase; gene Gene expression analysis in molecular biology experiments, the more commonly used methods, mainly Northern blot and dot blot hybridization (dot blot) and so on, without the need for dot blot transfer membranes, and the DNA or RNA is relatively fixed in a certain area, the requi