It evolved from Southern blotting hybrid method
Blotting hybridization Southern blot hybridization: gel ionizes DNA fragments digested with restriction enzymes, denatures the DNA on the gel and transfers single-stranded DNA fragments to a nitrocellulose membrane or other solid On the phase support, it is dried and fixed, and then reacted with the labeled probe of the corresponding structure at that time, and the color is developed by autoradiography or enzyme reaction, and the content of molecules of a specific size is detected. It can be used to analyze the restriction map of cloned genes, qualitative and quantitative analysis of genomic genes, gene mutation analysis and restriction length polymorphism analysis (RELP).
Northern blot hybridization: It evolved from Southern blotting hybrid method, and the tested sample is RNA. After formaldehyde or polyglyoxal denaturation and electrophoretic separation, it is transferred to a solid support and subjected to a hybridization reaction to identify the amount and size of a specific mRNA molecule in the group. This method is a commonly used method for studying gene expression, which can deduce the expression level of oncogenes.
Differential hybridization is the transfer of recombinant phage DNA from a genomic library to a nitrocellulose membrane, using two different types of cDNA probes (eg, reverse-transcribed cDNA from metastatic and non-metastatic cancer tissues) Hybridize to the DNA on the filter, and analyze the hybridization information at the corresponding positions on the two filters to isolate the differentially expressed genes.
It is suitable for china plastic tube factory of genes expressed by eukaryotes with less complex genomes (such as yeast), and the false positive rate is low. However, for very complicated genomes such as human beings, the percentage of expressed sequences is relatively low (only about 5%) due to the heavy workload, which is of little value. cDNA microarray hybridization refers to arranging cDNA clones or PCR products of cDNA in a high array and binding them to a solid support (such as a nylon membrane or an activated glass slide). Dot matrix, and then use a mixture of different DNA probes to hybridize the DNA on the microarray. Then, the fluorescence information, chemiluminescence, and confocal microscopy are used to scan the hybridization information on the microarray. It is more efficient, faster and cheaper than differential hybridization, and is suitable for large-scale analysis.
