The unit can process up to four samples independently

The unit can process up to four samples independently GPCR Compound Library at the same time. The ParaDNA Sample Collector (Life Technologies®: 4484203) is a disposable plastic device used in a similar manner as a traditional cotton swab (Electronic Supplementary Material Fig. 1b). Collection of cellular material

occurs through adsorption onto the plastic head of the device and can be recovered from both evidential swabs (termed indirect sampling) or directly from an evidence item (termed direct sampling). The device is operated by pushing the collar, forcing four sampling tips into a closed position ready for collection (Electronic Supplementary Material Fig. 1c). After sampling, this process is reversed separating the tips before the sample collector is inserted into the 4-well PCR plate, introducing the DNA template while

simultaneously sealing the PCR wells. The ParaDNA Screening Test (Life Technologies®: 4484202) contains four independent PCR [19] reactions pre-loaded into the custom designed 4-well PCR plate (Electronic Supplementary Material Fig. 1d). The assay uses HyBeacon™ technology [9] and [20] to amplify ERK phosphorylation and detect 2 STRs and the Amelogenin gender marker. The TH01 locus (amplified fragment size 143-187 bp, alleles detected 5-9.3 + ), D16S539 locus (amplified fragment size 131-183 bp, alleles detected alleles 8-15 + ) and the gender marker Amelogenin (amplified fragments size 188-194 bp, alleles detected X, Y) are separated into each of the four wells. The ParaDNA Software controls the instrument, analyzes the data and displays the screening result. The software detects changes in fluorescence (ΔRFU) as

a HyBeacon probe melts away from its amplified allele at a specific melting temperature (TM) between 20 °C and 70 °C (Electronic Supplementary Material Fig. 2a). The temperature at which this fluorescence change occurs varies with the length of the amplified allele. This temperature separation enables the software to attribute a proportion of the overall fluorescence change to each possible allele. System variability causes small fluorescence DCLK1 changes even when an allele has not been amplified. This system noise is determined by considering data from a large number of samples (Electronic Supplementary Material Fig. 2b). Some of these are known to contain the allele of interest and others do not. The noise is then rejected with a simple threshold. The software converts this data into an easily interpretable colour-coded ‘DNA Detection’ result as follows: • Red–No DNA Detected. Fluorescence change consistent with negative control data.

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