Dot Blots

Aliquots of RNA were denatured by heating to 60℃in 7.5%formaldehyde，10 mmol/L Na-phosphate（pH 6.8），for 30 min，according to the procedure of White and Bancroft［30］.For analysis by hybridization with radioactive DNA probes，series of RNA containing dots were formed on strips of Gene Screen Hybridization Transfer membrane（New England Nuclear）.Each 5-mm diameter dot was formed by applying 5 μg RNA in 10 μl to the Gene Screen Membrane presoaked in 2XSSC（1XSSC=0.15 mol/L NaCl，0.015 mol/L Nacitrate）by gentle capillary action using a stack of four layers of wet 3 MM-Whatman paper and dry paper toweling（about 2 cm high）underneath the Gene Screen.Dot blots were airdried and membranes were baked in a vacuum at 80℃for 4 h.(https://www.daowen.com)