Abstract: The invention discloses means and methods for genotyping an individual head of cattle. Individual's DNA is genotyped utilising the herein defined PCR SNaPshot protocol. The protocol comprises two PCR steps where the first step (PCR1) includes adding primers (SEQ ID No. 1-30) and/or primers extended at their 5? end with a common 10 base motif (ACGTTGGATG) to the PCR1 reaction. The second step (PCR2) includes adding extension primers (SEQ ID No. 31-45), and/or primers adjacent to corresponding specific SNPs. Further steps include producing amplicons from a PCR1 mixture comprising template DNA and the first primer set to yield PCR1 products, using PCR1 products as templates to a set of extension primers to yield PCR2 products. Size and colour separation is achieved by adding tails of different lengths to the PCR2 primers. PCR2 products are separated and the results compared with SNP profiles from the databank to obtain matching.
Type:
Application
Filed:
August 4, 2016
Publication date:
December 15, 2016
Applicants:
BACTOCHEM LTD., STATE OF ISRAEL, MINISTRY OF AGRICULTURE AND RURAL DEVELOPMENT, A.R.O. -- VOLCANI CENTER
Inventors:
Aviv CAHANA, Andrey SHIRAK, Baruch KARNIOL, Yitzchak SKALSKY, Joel Ira WELLER, Micha RON, Eyal SEROUSSI
Abstract: The invention discloses means and methods for genotyping an individual head of cattle. Individual's DNA is genotyped utilizing the herein defined PCR SNaP-shot protocol. The protocol comprises two PCR steps where the first step (PCR1) includes adding primers (SEQ ID No. 1-30) and/or primers extended at their 5? end with a common 10 base motif (ACGTTGGATG) to the PCR1 reaction. The second step (PCR2) includes adding extension primers (SEQ ID No. 31-45), and/or primers adjacent to corresponding specific SNPs. Further steps include producing amplicons from a PCR1 mixture comprising template DNA and the first primer set to yield PCR1 products, using PCR1 products as templates to a set of extension primers to yield PCR2 products. Size and color separation is achieved by adding tails of different lengths to the PCR2 primers. PCR2 products are separated and the results compared with SNP profiles from the databank to obtain matching.
Type:
Grant
Filed:
June 14, 2009
Date of Patent:
September 13, 2016
Assignees:
BACTOCHEM LTD., STATE OF ISRAEL, MINISTRY OF AGRICULTURE AND RURAL DEVELOPMENT, A.R.O.—VOLCANI CENTER
Inventors:
Aviv Cahana, Andrey Shirak, Baruch Karniol, Yitzchak Skalsky, Joel Ira Weller, Micha Ron, Eyal Seroussi
Abstract: The invention discloses means and methods for genotyping an individual head of cattle. Individual's DNA is genotyped utilising the herein defined PCR SNaP-shot protocol. The protocol comprises two PCR steps where the first step (PCR1) includes adding primers (SEQ ID No. 1-30) and/or primers extended at their 5? end with a common 10 base motif (ACGTTGGATG) to the PCR1 reaction. The second step (PCR2) includes adding extension primers (SEQ ID No. 31-45), and/or primers adjacent to corresponding specific SNPs. Further steps include producing amplicons from a PCR1 mixture comprising template DNA and the first primer set to yield PCR1 products, using PCR1 products as templates to a set of extension primers to yield PCR2 products. Size and colour separation is achieved by adding tails of different lengths to the PCR2 primers. PCR2 products are separated and the results compared with SNP profiles from the databank to obtain matching.
Type:
Application
Filed:
June 14, 2009
Publication date:
April 21, 2011
Applicant:
BACTOCHEM LTD.
Inventors:
Aviv Cahana, Andrey Shirak, Baruch Karniol, Yitzchak Skalsky, Joel Ira Weller, Micha Ron, Eyal Seroussi