# Mutation Rate Model Used in the DNA VIEW Program
## 2 Materials and Methods  
All computations were performed using large database of allelic DNA frequencies obtained from the Medical University of Lublin. This database contains DNA results of 7076 individuals, including personal information such as names, dates, places of birth etc. From this database we took 4410 unrelated persons and computed the distribution of allele frequencies of this sample (for example Table 1 presents the frequencies for the D5S818 locus). Three-thousand-nine-hundred-and-seventy-three individuals were typed using the classical electrophoresis method and 437 by NGS (next generation sequencing). All individuals were genotyped within 17 STR (short tandem repeat) markers located on the autosomal chromosomes. The techniques of DNA amplification are well described in [PERSON]'s book [23] in Section 1. The loci of 3973 individuals were amplified using commercial kits: PowerPlex 16 Hs System and PowerPlex 17 ESX System (Promega Corporation, Madison, WI, USA). PCR products were separated and detected on the ABI 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). Data were analyzed with the Gene Mapper ID software (v3.2, Applied Biosystems, Foster City, CA, USA, 2005). Laser-induced fluorescence was used in CE systems (Capillary Electrophoresis) with detection (by sensors) as low as \(10^{-18}\) to \(10^{-21}\) mol. The sensitivity of the techniques is attributed to the high intensity of incident light and the ability to accurately focus the light on the capillary. Multi-color fluorescence detection can be achieved by including multiple dichroic mirrors and bandpass filters to separate the fluorescence emission among multiple detectors (e.g., photomultiplier tubes) or by using a prism or grating to project spectrally resolved fluorescence emission onto a position-sensitive detector (sensor) such as a CCD (Charge Coupled Device) array. CE systems with 4- and 5-color LIF (Laser-Induced Fluorescence) detection systems are used routinely for capillary DNA sequencing and genotyping (DNA fingerprint) applications. The 437 individuals were profiled by next-generation sequences (NGS). Libraries were prepared using the ForenSeq DNA Signature Prep. Kit (Illumina) according to the manufacturer's protocol. Sequencing was performed on MiSeq FGx using the Miseq FGx Reagent Kit v.3 (600 cycles) and the ForenSeq Universal Analysis Software package. Following cluster generation, clusters were imaged using LED and filter combinations specific to each of four fluorescently labeled dideoxynucleotides. After the imaging of a tile was complete, the flow cell was moved into place to expose the nest \begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline \(i\) & \(n_{i}\) & \(v_{i}\) & \(i\) & \(n_{i}\) & \(v_{i}\) & \(i\) & \(n_{i}\) & \(v_{i}\) \\ \hline
1 & 9 & 0.00022 & 7 & 15 & 0.15693 & 13 & 20 & 0.02359 \\
2 & 10 & 0.00730 & 8 & 16 & 0.17187 & 14 & 21 & 0.00999 \\
3 & 11 & 0.01404 & 9 & 17 & 0.13188 & 15 & 22 & 0.00550 \\
4 & 12 & 0.09717 & 10 & 17.3 & 0.00011 & 16 & 23 & 0.00157 \\
5 & 13 & 0.09998 & 11 & 18 & 0.08324 & 17 & 24 & 0.00056 \\
6 & 14 & 0.15850 & 12 & 19 & 0.03752 & & \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Allele (indicated by the number of gene repetition); \(n_{i}\) frequencies; \(v_{i}\) at the D5S818 locus \(N=17\), \(\omega_{M}=0.00025,\omega_{F}=0.0012\), \(\omega=0.0011\) (cf. [24]). tile. The process was repeated for each cycle of sequencing. Following image analysis, the software performed base calling, filtering, and quality scoring. All amplification results were stored in a database. All computations were performed in the R-3.6.0 language on a RSTUDIO platform (v1.2.1335, RStudio, Boston, MA, USA, 2019). The starting point for our considerations is the following allele frequencies table (for example, for locus \(D55818\)).