Analysis of fast recognition in crops to determine the insertion of donor

摘要

Claim 1: a method to detect the specific Integration site Sequence polinucleu00f3tido Donor site within a genomic Target, wherein the method comprises: a) to amplify Genomic DNA with a first round of PCR to produce a First amplicon,Where the first PCR was performed using a PCR primer output designed to bind to the target Site of genomic PCR primer and a First input designed to bind to the sequence of polinucleu00f3tido Donor; (b) amplify the first amplicon with a second round of PCR or Using Specific primers sequences located within the first amplicon.To produce a Second amplicon; and (c) detecting the presence of a Second amplicon, where production of a Second amplicon indicates the presence of a specific event of Integration site.Claim 10: The Method according to claim 1, wherein the Genomic DNA that includes the specific Integration site Sequence polinucleu00f3tido Donor site within the genomic target is a Plant Genomic DNA.Claim 18: a method to detect the specific Integration site Sequence polinucleu00f3tido Donor site within a genomic Target plant cells transfected, wherein Said method comprises (a) an Amplify Genomic DNA with a first round of PC R to produce a First amplicon,Where such PCR is performed using a First output PCR primer designed to bind to the target Site of genomic PCR primer and a First input designed to bind to the sequence of donor polinucleu00f3tido,In addition, where the first PCR primer entrance is provided at a Lower concentration than the first PCR primer to amplify output; (b) the first amplicon with a second round of PCR using Sequence specific primers located within the first amplicon to pro While a Second amplicon; and (c) detecting the presence of a Second amplicon,Where the production of a Second amplicon indicates the presence of a specific event of Integration site.