INTRODUCTION
In current time, the rate of death caused by bleeding is increasing and there are limitations on how to resolve the medical problem instantly. Consequently, the group is planning to use the genes that allow lizards to regenerate/heal faster to improve the veti-gel (anti-bleeding gel). This modification and innovation will allow the veti-gel not only to stop bleeding, but also to hasten the process of cell healing.
REVIEW OF RELATED LITERATURE
McLean et al., 2011
- Regeneration is the process of renewal, restoration, and growth by which some organisms replace lost body parts and a number of regenerative mechanisms which is being evolved by different species
- Regeneration may produce genomes, cells, organisms, and ecosystems resilient. Regeneration is also most common in invertebrates such as the leopard gecko wherein a tail which is detached may regenerate naturally. The reason behind this is because the tail of a gecko represents roughly 41% of total body length and is composed of multiple tissue types including striated muscle, a bony vertebral column, spinal cord and more.
- The difference between lizards and geckos is that geckos retain the notochord into skeletal maturity.
McLean et al., 2011
Regarding the type of regeneration done by leopard geckos, it is called Epimorphic regeneration in which Blastema, or the restoration of lost tissues and structures from an aggregation of proliferating cells is involved.
PROBLEM
In our modern society, accidents, whether it be big or small, can happen anytime. According to the World Health Organization (WHO), the number of deaths caused by traffic accidents here in the Philippines was around one million back in 2011. These deaths may be caused due to excessive bleeding after the accidents or the late arrival of the patient to the hospital. Just imagine if paramedics have the power to stop the bleeding as soon as possible and instigate healing of the wound as quickly as possible.
OBJECTIVE
- to create a gel that could stop the bleeding of a wound or a cut, and heal it as quickly as possible.
- to find out which gene of the lizard helps cells reproduce faster to be able to regenerate loss tissues
METHODOLOGY
MATERIALS:
1. 2.5 cm square Lizard Skin
2. Sterile scalpel blade
3. 2.5 mL Eppendorf tubes
4. 900 µL of lysis buffer
5. 4 µL of RNAse A
6. 300 µL of 7.5 M ammonium acetate
7. Laboratory centrifuge
8. 900 µL of isopropanol
9. 500 µL of 70% ethanol
10. 30-100 µL of TE Buffer
11. Ice
PROCEDURE
7. To suspend the DNA again, add substance in 30-100 µL of TE Buffer, depending on the size of the pellet.
8. When not in use, store DNA at -20°C.
9. After extracting the DNA, inject DNA substance into the Veti-Gel.
PROCEDURE
5. To precipitate the DNA, transfer the supernatant to a new tube containing 900 µL of isopropanol and invert tube 20 times to mix and pellet the DNA. After this, centrifuge it to pellet the DNA.
6. To wash the DNA, pour off the isopropanol and wash the pellet in 500 µL of 70% ethanol. Centrifuge to pellet the DNA, remove the ethanol and allow residual ethanol to evaporate from the inverted tube.
1. Cut the 2.5 cm square Lizard skin into small pieces using a sterile scalpel blade.
2. Add the skin bits to the 900 µL of lysis buffer and 9 µL of proteinaseK. Heat it at 55°C for 5-12 hours.
3. To remove the RNA, cool the substance to room temperature and add 4 µL of RNAse A. Incubate at 37°C for one hour.
4. Now to extract the DNA, cool the substance to room temperature and add 300 µL of 7.5 M ammonium acetate. Mix and stand in ice for 1-15 minutes. Centrifuge at top speed (13k RPM) for 3 minutes to pellet the skin and cell debris.
Cho•Hernandez
Reyes•See•Wong•Wu
IMPLICATION/SIGNIFICANCE
- This biotechnology project would be implied, but not limited to the field of medicine.
- Death rate due to bleeding would significantly decrease once further study about this experiment would be conducted.
- Lizard veti-gel, an anti-bleeding gel would serve as an innovative and powerful biotechnological project that would greatly be beneficial to patients suffering from different illnesses, most especially those suffering from internal bleeding.
- The group’s project, the lizard veti-gel is unique because it will greatly help in a low cost with effects that are yet to exist in the society.
- This gel would not only stop bleeding but also hasten the process of cell healing.
- The group carefully selected lizards because lizards regenerate and have the ability to mimic some cells of the human body to make the healing process faster.