cre PCR Sample Clauses

cre PCR. A PCR assay for cre was used to quantify donor DNA in recipient DLN of mice receiving either wild-type or ephrin B2-/- heart grafts; as the donor mice had been transduced with the cre gene, and therefore DPL within the grafts contained a copy of the gene. PCR reactions were made up to a final volume of 20µl in white hard-shell 96-well PCR plates (Bio-Rad Laboratories, Inc., Hercules, CA, USA). PCR reactions consisted of: 10µl Taqman (Invitrogen; Thermo Xxxxxx Scientific, Inc.), 1µl primer mix (assay Mr00635245, FAM conjugated probe); (Applied Biosystems; Thermo Xxxxxx Scientific, Inc.), and genomic DNA (extracted from DLN as above) diluted to a volume of 9µl in DNase free water (Qiagen, Inc.). After addition of reaction mixtures to xxxxx of the plate, the plate was spun for 1 minute at 1500RPM. The PCR conditions were as follows: an initial hold step of 50ºC, then heating to 95ºC, and 40 cycles of 15 seconds at 95ºC followed by annealing for 1 minute at 60ºC. Data was analysed using the Bio-Rad CFX software.
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