The Optimization of Southern Blot for Transgenic Sugarcane Plants

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.015

Abstract

Abstract: Using sugarcane as materials, the DIG-labeled Southern blot was optimized through several key aspects:the comparison of different-labeled probe methods, extraction of sugarcane genome DNA, the amount of enzyme digestion of genome DNA, enzyme digestion time and a serial of procedures in the process of the hybrid. The results showed that sugarcane DNA extracted by the improved CTAB method met the requirements of the latter experiments, the efficiency of PCR-labeled probe was higher than that of random primer-labeled probe, so PCR-labeled probe method was suitable for the Southern blot, 40 μg DNA samples in enzyme digestion system of 400 μL for 10 hours could achieved desirable digestion result；while hybrid temperature was 40℃ and hybridizing time was 18 h, a clear hybridization band could be observed. The optimization of sugarcane Southern blot provides a reference for the analysis of Southern blot of transgenic sugarcane.

Key words: sugarcane, Southern blot, digoxigenin, PCR-labeled probe, enzyme digestion

Cui Xueqiang, Zhang Shuzhen, Shen Linbo, Feng Cuilian. The Optimization of Southern Blot for Transgenic Sugarcane Plants[J]. Biotechnology Bulletin, 2015, 31(12): 105-109.

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