Calculation of cell number Sample Clauses

Calculation of cell number. The CFX software calculates the amount of target DNA (i.e. zfy1 or cre) in samples by comparing ct values to a standard curve. As the copy number of the cre transgene in donor mice was unknown, individual standard curves using DNA from each donor mouse were run at the same time as the recipient DLN DNA samples. For the zfy1 assay this was not necessary as all male donor mice have one copy of the gene per cell (214). In order to calculate the number of donor cells within recipient DLN DNA samples from the PCR assays the c value was used. The c value denotes the size of an organism’s genome in terms of the mass of DNA (in picograms) in a haploid cell, i.e. a gamete, or half the amount of DNA in a diploid cell. The c-value for mus musculus is 2.45 (216). Therefore, the number of donor cells was determined by calculating the percentage of the total sample DNA which was donor specific (i.e. the proportion value), converting this to a concentration (picograms of DNA) and dividing by 4.9 (double the c-value).
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