Investigating mutation of human ?-globin gene by PCR

Extract

[...] Figure Annotated Wild Type Sequence As seen in Figure the 3 exons and 2 introns have been annotated on the gene sequence. The exons are labelled "exon and and the introns are labelled "intron 1 and 2". Exons can also be identified by light green, light grey and dark grey. Introns can also be identified by the colours orange and purple. Figure Identifying Mutation Site in Wild Type Sequence As seen in Figure the mutation site was identified and annotated in the wild-type sequence. This can be seen by the colour pink which marks the mutation site. [...]

[...] In Micromachines (Vol Issue 5). https://doi.org/10.3390/mi12050519 2. Laboratories, B.-R. (2014). PCR Troubleshooting. Life Science Research, 800. 3. Mangla, A., Ehsan, M., Agarwal, N., & Maruvada, S. (2023). Sickle Cell Anemia. StatPearls [Internet]. https://www.ncbi.nlm.nih.gov/books/NBK482164/#:~:text=Sickle%20cell%20anemia%20is%20an,pain%20crises%2C%20and%20organ%20damage. 4. [...]

[...] Needs, T., F. Gonzalez-Mosquera, L., & T. Lynch, D. (2023). Beta Thalassemia. StatPearls [Internet]. https://www.ncbi.nlm.nih.gov/books/NBK531481/ 5. Wu, S. S., Li, Q. C., Yin, C. Q., Xue, W., & Song, C. Q. (2020). [...]

[...] The mutation site annotated contains the base pairs "CCTGAG" since it is the wild type. Figure Mutant Sequence With Mutation Site As seen in Figure a mutant-type sequence is generated from the wild-type sequence. This was done by replacing the adenine in "CCTGAG" from the wild-type sequence with thymine in the mutant sequence to give "CCTGTG". The wet lab experiments were performed according to the protocol provided in the lab. Results: Figure Primer Design As seen in Figure the forward and reverse primers were designed for the wild-type sequence. [...]

[...] Investigating mutation of human ?-globin gene by PCR Abstract: This comprehensive study investigates the molecular basis of sickle cell anaemia (SCA) by employing bioinformatics tools and PCR techniques to analyze mutations in the HBB gene. The research effectively differentiates between wild-type and mutant gene sequences by designing specific primers. The study confirms the accuracy of PCR in detecting SCA, aligning laboratory results with in-silico predictions. Additionally, it discusses the potential of advanced gene editing methods like CRISPR-Cas9 for future therapeutic approaches and emphasizes the importance of longitudinal studies for a deeper understanding of disease progression and the impact of various mutations on individuals with sickle cell traits. [...]