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?
nuDNA typing system, InnoTyper 21.
It is a small amplicon retrotransposon marker
typing system.
A bi-allelic marker system that contains 20 Alu
element markers, ranging in size from 60 to 125
bp.
Able to retrieve DNA from highly degraded,
compromised and with little sample of rootless
hair shaft.
Advantages of Innotyper21?
1. Less labor-intensive compared to mtDNA
test
2. High sensitivity. (Works well with small
sample size)
3. Bi-allelic system simplifies data analysis
4. It has the smallest amplicon size range,
providing an advantage for extremely
challenging forensic samples.
With Innotyper21 it will increase the chance of
retrieving DNA sample from hair, thus, solving
the crime.
 Reviewing of article…..
Sample preparation/DNA Extraction
- Hair follicular tag is identified with
stereomicroscopy and removed with
UV-irradiated scissors and cleaned with
sonication.
- Samples are digested in Buffer ATL
with Proteinase K and 1M DTT,
incubated for 1hr @ 56℃, followed by
addition of Buffer AL and further
incubation for 10mins @ 70℃.
- DNA is purified using BTA Forensic
DNA extraction kit which uses magnetic -
beads to isolate DNA. Finally DNA is
eluted in PrepFiler Elution Buffer.
DNA Quantification/Amplification
- Purified DNA are quantified with
InnoQuant®, to determine the
degradation index of the
samples.
- Samples are amplified with
InnoTyper 21 system with
modified reaction volumes and
PCR cycles.
- InnoTyper 21 uses small
amplicons for amplification.
Retrotransposable Insertion
Polymorphism (RIP) markers are
included.
DNA
Sequencing/Analysis
- AB 3130 Cpillary
Electrophoresis
Genetic Analyzer
and Southern
separates and detect
amplified fragments.
- Results are analysed
with GeneMapper®
ID-X using the Local
Southern size calling
method.
 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].