Sequence analysis

DNA sequencing of the 18S rDNA fragment

Purification of PCR product

Analysis of internal transcribed spacer region

RAPDand SSRscoring and data analysis

PCR amplification

Running of gel and visualization of DNA

Determination of the yield

Agarose gel electrophoresis

Qualitative and quantitative estimation of DNA

Determination of the yield

Procedure for DNA isolation

Reagents required for fungal DNA isolationand p

Last node to spike length(cm)

Endomorphic ectomorph:

SOMATOTYPING

Lysis Buffer: 0.1% Triton X-100/0.1 M Tris-HCl (pH 8.0). 450 ml distilled water 50 ml 1M Tris-HCl (pH 8.0) 0.5 ml Triton X-100 detergent • 100X Mg++ solution: 0.1 M magnesium chloride 4.5 M 2-mercaptoethanol Stored at 4°C. • 0.1 M sodium phosphate (pH 7.5)41 ml 0.2 M Na2HPO4 9 ml 0.2 M Na H2PO4 50 ml distilled water • 4 mg/ml ONPG (o-nitrophenyl-β-D-galactopyranoside) in 0.1 M sodium phosphate (pH 7.5) containing 2 mM β-mercaptoethanol, Stored at –20°C. • 0.1 mg/ml β-gal standard: 0.1 mg/ml β-gal in 0.1 M sodium phosphate (pH 7.5) containing 2 mM 2-mercaptoethanol Stored at 4°C. • 1 M sodium carbonate in water

Solutions: