Noninvasive method of DNA isolation from fecal epithelial tissue of dairy animals.
A novel noninvasive genomic DNA isolation protocol from fecal tissue, by the proteinase K digestion and guanidine hydrochloride extraction method, was assessed for the genotyping of cattle and buffalo. The epithelial tissues present on the surface of the feces were used as source for isolation of genomic DNA. The DNA isolated from fecal tissue was found to be similar as those obtained from other body tissues such as skin, brain, liver, kidney, and muscle. The quality of DNA was checked by agarose gel electrophoresis and polymerase chain reaction (PCR). We successfully amplified a 320 bp MHC class II DRB gene and a 125 bp mt-DNA D-loop region from isolated genomic DNA of cattle. Thus, the DNA isolated using this method was suitable for common molecular biology methods, such as restriction enzyme digestion and genotyping of dairy animals through PCR.
a National Dairy Research Institute , Animal Biotechnology Centre , Karnal , India.
Bidhan
B
Chandra De
Mahesh Chandra
MC
Patra
Sushil
S
Kumar
Biswajit
B
Brahma
Devika
D
Goutam
Latika
L
Jaiswal
Ashutosh
A
Sharma
Sachinandan
S
De
D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D004847:Epithelial Cells / Q000737:chemistry*; D005243:Feces / Q000166:cytology*; D005260:Female; D016133:Polymerase Chain Reaction
D004247:DNA
Bovine; DNA isolation; Fecal epithelial tissue; Noninvasive