Oligonucleotide probe developed by using specific sequence of elytrigia elongata
A technology of E. elongatum and oligonucleotides, applied in the field of cell biology, can solve problems such as the limitation of chromosome karyotype, and achieve the effects of abundant positions, stable hybridization signals, and accurate identification
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[0028] In order to verify the feasibility of the oligonucleotide probe Oligo-pTh1 and clarify its fluorescence signal intensity, the combination of Oligo-pTh1 and Oligo-pTh2 was used to conduct fluorescence in situ hybridization experiments in the Chinese spring-Thinopyrum elongatum addition line. The specific operations are as follows:
[0029] (1) Send the sequences of Oligo-pTh1 and Oligo-pTh2 to the company for synthesis, and modify the 5' ends of the sequences with TAMRA and FAM respectively;
[0030] (2) The synthesized probe is diluted according to the concentration requirements, and the hybridization solution is prepared according to the system in Table 1. The prepared hybridization solution is denatured in a metal bath at 103°C for 13 minutes. In cold alcohol, place it in a refrigerator above -20°C;
[0031] (3) Put the prepared chromosome sheet of the Chinese spring-Echinopsis elongatum addition line in 70% alcohol solution of 0.15mol / L NaOH for denaturation for 5min...
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