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Nuclear DNA profiling on rootless

hair shaft (Innotyper 21)


Team Members:
Lim Zheng Yong (19113698)
Lim Rui An (19155403)
Celeste Ong (19546782)
Amelia Lee (19551292)
Aisha Rashid (19120407)
Reasons we chose this topic….
Hair evidence can be substantial in solving crimes as it put a victim and suspect together at a
crime scene. Hairs can be transferred through direct or indirect means.

Forensic scientists can conclude several important informations from analyzing hairs found at
crime scenes. Some clues that hair reveals include:

- part of body it came from

- race of person

- hair shed or pulled from someone's head

- any color treatment.


Introduction….

Researched on?
Advantages of Innotyper21?
nuDNA typing system, InnoTyper 21.
1. Less labor-intensive compared to mtDNA
It is a small amplicon retrotransposon marker test
typing system. 2. High sensitivity. (Works well with small
sample size)
A bi-allelic marker system that contains 20 Alu 3. Bi-allelic system simplifies data analysis
element markers, ranging in size from 60 to 125 4. It has the smallest amplicon size range,
bp. providing an advantage for extremely
challenging forensic samples.
Able to retrieve DNA from highly degraded,
compromised and with little sample of rootless With Innotyper21 it will increase the chance of
hair shaft. retrieving DNA sample from hair, thus, solving
the crime.
Reviewing of article…..
DNA
Sample preparation/DNA Extraction DNA Quantification/Amplification
Sequencing/Analysis

- Purified DNA are quantified with


- Hair follicular tag is identified with
InnoQuant®, to determine the - AB 3130 Cpillary
stereomicroscopy and removed with
degradation index of the Electrophoresis
UV-irradiated scissors and cleaned with
samples. Genetic Analyzer
sonication.
- Samples are digested in Buffer ATL - Samples are amplified with and Southern
with Proteinase K and 1M DTT, InnoTyper 21 system with separates and detect
incubated for 1hr @ 56℃, followed by modified reaction volumes and amplified fragments.
addition of Buffer AL and further PCR cycles. - Results are analysed
incubation for 10mins @ 70℃. - InnoTyper 21 uses small with GeneMapper®
- DNA is purified using BTA Forensic amplicons for amplification. ID-X using the Local
DNA extraction kit which uses magnetic - Retrotransposable Insertion Southern size calling
beads to isolate DNA. Finally DNA is Polymorphism (RIP) markers are method.
eluted in PrepFiler Elution Buffer.
included.
Critical Analysis (Discussion).....
Extracted DNA from individual samples were variable

- The sample collected were either able to yield or not able to yield a useable profile.

Degradation indices ranging from 2 to 114 with an average of 33

- Indicating highly degraded of nuDNA

Factors influencing allele recovery

- Extent of DNA degradation


- Amount of DNA inserted for amplification
- Cosmetic use will influence quantification of DNA
- Quality control when handling and testing hair sample.
In Conclusion…..

- 40% of the samples were interpretable despite having low amount and
degraded DNA
- Higher discrimination power and statistical power of Innotyper 21 raises
possibility of replacing mtDNA in examining hair shafts
- Increased efficacy in processing hair shafts in combination with current PCR
assays

Future explorations

- Devise an improved and standardized protocol to reduce variable results


- Quality control
- Washing technique for the hair shafts
References….
1. Grisedale, K.S., Murphy, G.M., Brown, H. et al. Int J Legal Med (2018) 132: 107.
https://doi-org.ez.library.latrobe.edu.au/10.1007/s00414-017-1698-z

2. Career Trend. (2018). The Importance of Hair Evidence in Forensics. [online] Available
at: https://careertrend.com/about-5445024-importance-hair-evidence-forensics.html
[Accessed 24 Jan. 2018].

3. Innogenomics.com. (2018). InnoTyper® 21 | InnoGenomics Technologies, LLC. [online]


Available at: http://innogenomics.com/products/innotyper-21/ [Accessed 24 Jan. 2018].

4. American Academy of Forensic Sciences. (2017) New Orleans, LA. Available at:
https://www.aafs.org/wp-content/uploads/2017Proceedings.pdf [Accessed 24 Jan.
2018].

5. GeneMapper® ID-X Software Version 1.0.1/1.1. (2009). [ebook] Applied Biosystems.


Available at: https://tools.thermofisher.com/content/sfs/manuals/cms_048198.pdf
[Accessed 24 Jan. 2018].