金属离子对总RNA片段化的影响

n important consideration in microarray analysis of nucleic acids is the efficiency with which the target molecule is captured by, or hybridized to, surface-immobilized nucleotide probe .As for RNA, secondary or tertiary structure of the target strand can significantly decrease the capture efficiency. To overcome this limitation, RNA is often fragmented to reduce the structural effects. The total RNA can be fragmented by using Divalent Ions and Alkali. The fragmentation of total RNA between 20~200 nt can be obtained with 0.1 mol /L Mg2+ at 95 ℃ for 15min or Ambion fragmentation reagent at 70 ℃ for 15 min or 0.1 mol /L NaOH at 37 ℃ for 20 min. The fragmentation of total RNA between 20~60 nt can be obtained with 0.1 mol /L Zn2+ at 95 ℃ for 15 min or 0.1 mol /L NaOH at 47 ℃for 20 min. This method can provides enough fragmented RNA quickly to hybridize with one genome-scale Gene Chip array. This study provides a new method of decrease the effect of RNA complicated structure for microarray chip.