Familial acute myeloid leukemia Screening of differentially expressed genes.docVIP

 PAGE \* MERGEFORMAT 7 Familial acute myeloid leukemia Screening of differentially expressed genes Author: Wang Shao-yuan, ZHANG Yi-wen, Wang Yi Cheng ‘ [Abstract] Objective To establish a simple and reliable non-isotopic hybridization screening methods, screening of familial acute myeloid leukemia subtractive cDNA library of differentially expressed genes. Methods digoxigenin (DIG) tag subtractive library of cDNA as a probe, through the differential screening technology for the screening of differentially expressed fragments, hybridization positive results by RT  PCR re-authentication. The results of DIG-labeled probe concentration of 250 ~ 500 pg / μ L, the efficiency is 48% ~ 96%, and the hybridization signals clear and reproducible. Application of DIG-labeled probe differences between the improved screening of positive results obtained by RT  PCR validation in line with rate of 86%. Conclusion The DIG-labeled probe with a high degree of sensitivity and good specificity of radioactive contamination can be avoided, simplifying experimental operation, can be used as an alternative isotope 32P  dCTP-labeled probe method. [Keywords:] pedigree, leukemia; acute; gene expression; digoxin; DNA, Complementary; gene library By suppression subtractive hybridization (suppression subtractive hybridization, SSH) technology to build a cDNA subtractive library, contains a wealth of information on differential expression is to study the molecular mechanism of disease development in an important source of information. However, how the information from a vast library was screened differentially expressed genes out of a valuable, requires a simple, practical, fast and reliable screening technology, which not only a highly efficient screening efficiency, but also should have a high degree of sensitivity and specificity of , while operation can not be cumbersome, can be carried out in an ordinary lab. Therefore, I established digoxin (digoxin, DIG)-labeled probe imp