Infection of ileum mesothelium: Methods IHC (Part 2)

hybridizationIn situ hybridization

Dewaxed and dehydrated sections were digested with 100 ^L of pepsin (Carezyme II, Biocare Medical, USA) at 37°C for 5 min. Slides were rinsed in Tris-buffered saline, dehydrated in ethanol and air dried. Ten microlitres to 20 ^L of biotin-labelled HSV type I, HSV type II, CMV and adenovirus probes (ENZOLife Sciences Inc, USA) were placed on slides and cover slipped. The sections with probes were denatured at 95°C for 5 min and the slides incubated at 37°C overnight. After hybridization, slides were washed in Tris-buffered saline until the cover slips detached. Digoxigenin-labelled hybrids were then identifiable by the ENZO detection kit (ENZOLife Sciences Inc). The slides were then counterstained with hematoxylin and cover slipped. buy levaquin online

PCR

DNA was extracted from histological slide bowel tissue and amplified by 40 cycles (approximately 2.5 h) according to a standard protocol. Amplified DNA was analyzed by gel electrophoresis for expected size products. Appropriate DNA was then exposed to dot blot testing to confirm the correct DNA sequencing. Material was tested against CMV primers (HSV -P1 [5′] and HSV -P2 [3′]), and varicella zoster virus (VZV) primers (VZV-P1 [5′] and VZV-P2 [3′]).

 

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