Low Throughput Direct Cycle Sequencing of Polymerase Chain Reaction (PCR) Products
Book chapter
Nikolaos Tairis and Zouganelis, G. 2023. Low Throughput Direct Cycle Sequencing of Polymerase Chain Reaction (PCR) Products. in: Scarlett, Gary (ed.) DNA Manipulation and Analysis New York, NY Springer. pp. 195–211
| Authors | Nikolaos Tairis and Zouganelis, G. |
|---|---|
| Editors | Scarlett, Gary |
| Abstract | Polymerase Chain Reaction (PCR) products have been traditionally characterized by cloning and cycle sequencing. However, when quick sequencing data are required, the cloning step may be omitted and PCR products can be sequenced directly. We describe here a sequencing protocol that involves the gold standard Big Dye chemistry in a low throughput format using one of the latest sequencing platforms the ABI Seqstudio. Our cycle sequencing protocol follows the following steps: (1) purification of the PCR product with a spin column-based kit; (2) quality & quantity assessment of the PCR product with the use of spectrophotometry & gel electrophoresis; (3) setup and amplification of the cycle sequencing reaction; (4) Capillary Electrophoresis; (5) Sequence Data Analysis. |
| Keywords | Sanger sequencing; PCR; cloning and cycle sequencing |
| Page range | 195–211 |
| Year | 2023 |
| Book title | DNA Manipulation and Analysis |
| Publisher | Springer |
| Humana | |
| Place of publication | New York, NY |
| Series | Methods in Molecular Biology |
| ISBN | 9781071630037 |
| 9781071630044 | |
| ISSN | 1064-3745 |
| 1940-6029 | |
| Digital Object Identifier (DOI) | https://doi.org/10.1007/978-1-0716-3004-4_15 |
| Web address (URL) | http://dx.doi.org/10.1007/978-1-0716-3004-4_15 |
| Output status | Published |
| Publication dates | Mar 2023 |
| Publication process dates | |
| Deposited | 17 May 2023 |
| Journal | Methods in Molecular Biology |
Permalink -
https://repository.derby.ac.uk/item/9yv7y/low-throughput-direct-cycle-sequencing-of-polymerase-chain-reaction-pcr-products
51
total views0
total downloads5
views this month0
downloads this month
Export as
Related outputs
In-silico Codon Context and Synonymous Usage Analysis of Genes for Molecular Mechanisms Inducing Autophagy and Apoptosis with Reference to Neurodegenerative Disorders.
Khandia, R., Gurjar, P., Romashchenko, V., Al-Hussain, S. A., Alexiou, A., Zouganelis G. and Zaki, M.E.A. 2024. In-silico Codon Context and Synonymous Usage Analysis of Genes for Molecular Mechanisms Inducing Autophagy and Apoptosis with Reference to Neurodegenerative Disorders. Journal of Alzheimer's disease : JAD. 99 (3), pp. 927-939. https://doi.org/10.3233/jad-240158Compositional constraints and selection forces dictate codon usage in human bocavirus
Khandia, R.,, Singhal, S., Kumar, U., Shende, K., Pandey, M.K., Povetkin, S.N., Rzhepakovsky, I.V., Raut, A., Alexiou, A., Dhama, K., Zouganelis, G. and Khan. A. A. 2023. Compositional constraints and selection forces dictate codon usage in human bocavirus. Journal of King Saud University - Science. 35 (6), pp. 1-11. https://doi.org/10.1016/j.jksus.2023.102726The Efficient Activity of Glabridin and its Derivatives Against EGFR-mediated Inhibition of Breast Cancer
Marios Papadakis, Arabinda Ghosh, Debanjana Ghosh, Nobendu Mukerjee, Swastika Maitra, Padmashree Das, Abhijit Dey, Souty M.Z. Sharkawi, Georgios D. Zouganelis, Athanasios Alexiou, Somdatta Yashwant Chaudhari, Ritika Sharma, Sonali Arun Waghmare, Gaber El-saber Batiha and Zouganelis, G. 2023. The Efficient Activity of Glabridin and its Derivatives Against EGFR-mediated Inhibition of Breast Cancer. Current Medicinal Chemistry. https://doi.org/10.2174/0929867330666230303120942Nanomaterial-induced toxicity in pathophysiological models representative of individuals with pre-existing medical conditions
Sreejesh Sreedharan, Zouganelis, G., Samantha J Drake, Gyanendra Tripathi and Ali Kermanizadeh 2022. Nanomaterial-induced toxicity in pathophysiological models representative of individuals with pre-existing medical conditions. Journal of Toxicology and Environmental Health, Part B: Critical Reviews. pp. 1-27. https://doi.org/10.1080/10937404.2022.2153456The fractal viewpoint of tumors and nanoparticles.
Alexiou, A., Tsagkaris, C., Chatzichronis, S., Koulouris, A., Haranas, I., Gkigkitzis, I., Zouganelis, G., Mukerjee, N., Maitr, S., Jha, N. K., Batiha, G. and Amjad, M. 2022. The fractal viewpoint of tumors and nanoparticles. Current Medicinal Chemistry. 30 (3), pp. 356 - 370. https://doi.org/10.2174/0929867329666220801152347Crinum jagus (J. Thomps. Dandy): Antioxidant and protective properties as a medicinal plant on toluene-induced oxidative stress damages in liver and kidney of rats
Salihu, M., Batiha, G.E.S., Kasozi, K.I., Zouganelis, G., Sharkawi, S.M., Ahmed, E.I., Usman, I.M., Nalugo, H., Ochieng, J.J., Ssengendo, I., Okeniran, O.S., Pius, T., Kimanje, K. R., Kegoye, E.S., Kenganzi, R. and Ssempijjai, F. 2022. Crinum jagus (J. Thomps. Dandy): Antioxidant and protective properties as a medicinal plant on toluene-induced oxidative stress damages in liver and kidney of rats. Toxicology Reports. 9, pp. 699-712. https://doi.org/10.1016/j.toxrep.2022.03.026Development and validation of anti-human Alpha synuclein DNA aptamer using computer modelling techniques—an in silico study
Rock, M., Zouganelis, G., Belchior de Andrade, A., Drake, S., Alexiou, A., Albrakati, A., Bathia, G. E. S. and Illingworth, T. 2022. Development and validation of anti-human Alpha synuclein DNA aptamer using computer modelling techniques—an in silico study. Journal of Integrative Neuroscience. 21 (1), p. 5. https://doi.org/10.31083/j.jin2101005Characterization and phylogeny of fungi isolated from industrial wastewater using multiple genes
Ezeonuegbu, B.A., Abdullahi, M.D., Whong, C.M., Sohunago, J.W., Kassem, H.S., Yaro, C.A., Hetta, H.F., Mostafa-Hedeab, G., Zouganelis, G. and Batiha, G.E.S 2022. Characterization and phylogeny of fungi isolated from industrial wastewater using multiple genes. Scientific Reports. 12, pp. 1-11. https://doi.org/10.1038/s41598-022-05820-9Photoperiod-Dependent Expression of MicroRNA in Drosophila.
Pegoraro, M., Fishman, B., Zonato, V., Zouganelis, G., Francis, A., Kyriacou, C. P. and Tauber, E. 2022. Photoperiod-Dependent Expression of MicroRNA in Drosophila. International Journal of Molecular Sciences. 23 (9), pp. 1-14. https://doi.org/10.3390/ijms23094935The Inclusion of a Matrix Metalloproteinase-9 Responsive Sequence in Self-assembled Peptide-based Brain-Targeting Nanoparticles Improves the Efficiency of Nanoparticles Crossing the Blood-Brain Barrier at Elevated MMP-9 Levels
Islam, Yamir, Ehtezazi, Parinaz, Cashmore, Andrew, Marinsalda, Elena, Leach, Andrew G., Coxon, Christopher R., Fatokun, Amos A., Sexton, Darren W., Khan, Iftikhar, Downing, James, Pluchino, Stefano, Sivakumaran, Muttuswamy, Teixido, Meritxell, Ehtezazi, Touraj and Zouganelis, George 2020. The Inclusion of a Matrix Metalloproteinase-9 Responsive Sequence in Self-assembled Peptide-based Brain-Targeting Nanoparticles Improves the Efficiency of Nanoparticles Crossing the Blood-Brain Barrier at Elevated MMP-9 Levels. Journal of Pharmaceutical Science. https://doi.org/10.1016/j.xphs.2020.12.004