TY - JOUR
T1 - Milk as a Source of Deoxyribonucleic Acid and as a Substrate for the Polymerase Chain Reaction
AU - Lipkin, E.
AU - Shalom, Anne
AU - Khatib, H.
AU - Soller, M.
AU - Friedmann, A.
PY - 1993
Y1 - 1993
N2 - Somatic cells in milk were used as a source of DNA and as a substrate for the polymerase chain reaction. Successful DNA extraction required a minimum total of 17 × 106 cells in the milk sample. The DNA yield per cell was highly variable but comparable, on average, with the yield from peripheral blood leukocytes. In all cases, direct polymerase chain reaction on milk samples, using a variety of primer pairs, yielded amplified products of correct size that were identical to those obtained by polymerase chain reaction of purified DNA extracted from milk or blood. Milk and DNA extracted from milk served as substrates for direct sequencing of part of the bovine growth hormone gene. Milk samples with added preservative were stored over 200 d without effect on the polymerase chain reaction. Thus, milk can often substitute for blood as a source of DNA for Southern blot analysis and is a preferred substrate for the polymerase chain reaction. The technical convenience of milk as a source of DNA can be expected to increase the field of application of marker-based methods for genetic analysis and genetic improvement of economic traits in dairy cattle.
AB - Somatic cells in milk were used as a source of DNA and as a substrate for the polymerase chain reaction. Successful DNA extraction required a minimum total of 17 × 106 cells in the milk sample. The DNA yield per cell was highly variable but comparable, on average, with the yield from peripheral blood leukocytes. In all cases, direct polymerase chain reaction on milk samples, using a variety of primer pairs, yielded amplified products of correct size that were identical to those obtained by polymerase chain reaction of purified DNA extracted from milk or blood. Milk and DNA extracted from milk served as substrates for direct sequencing of part of the bovine growth hormone gene. Milk samples with added preservative were stored over 200 d without effect on the polymerase chain reaction. Thus, milk can often substitute for blood as a source of DNA for Southern blot analysis and is a preferred substrate for the polymerase chain reaction. The technical convenience of milk as a source of DNA can be expected to increase the field of application of marker-based methods for genetic analysis and genetic improvement of economic traits in dairy cattle.
KW - deoxyribonucleic acid extraction
KW - milk
KW - milk somatic cells
KW - PCR
KW - polymerase chain reaction
KW - polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=0027636296&partnerID=8YFLogxK
U2 - 10.3168/jds.S0022-0302(93)77536-0
DO - 10.3168/jds.S0022-0302(93)77536-0
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C2 - 8345129
AN - SCOPUS:0027636296
SN - 0022-0302
VL - 76
SP - 2025
EP - 2032
JO - Journal of Dairy Science
JF - Journal of Dairy Science
IS - 7
ER -