P-MEXP-1022 - P-MEXP-1022

Type
hybridization
Description
Catma reflective slides are pretreated for 30min with 2x SSPE/0.2%SDS at room temperature. They are dipped a couple of times in distilled water at RT and immediately dried by centrifugation. The slide was placed in a chamber with a 25x60 lifterslip and 13ul of 3x SSC for each groove. The probe was diluted to a final volume of 100 uL as follows: 9ul of purified, labeled cDNA; 6 ul of dH2O; 10 ul of Yeast tRNA (2ug/uL); 25 ul of Amersham Microarray Hybridization Buffer; 50 uL formamide. The probe mixture is heated for 3 min at 95oC, put on ice for 30 sec and centrifuged to remove dust for 1 min. The chamber (with the chip) and the probe mixture are preheated for 2-3 min at 42oC then added to the chip at quickly as possible. The microarray is sealed in a chamber and submerged in a 42oC water bath for approximately 16 h. The microarray is washed for 4 min in 1xSSC, 0.2% SDS (42oC); 4 min in 0.1x SSC, 0.2% SDS (RT); 4 min in 0.1x SSC, 0.2% SDS (RT); 4 min in 0.1x SSC(RT); and 4 min in 0.1x SSC(RT) then dried immediately by centrifugation (2min at 2600rpm)
Parameters
Chamber type, Quantity of label target used, Temperature, Time, Volume
Links
All experiments using protocol P-MEXP-1022: (E-CAGE-1, E-CAGE-2, E-CAGE-3, E-CAGE-4)