Expression of an intron-containing gene from P. falciparum using three sequential PCR steps
The amplification of eukaryotic genes, with introns is very tedious and expensive. The only available lab method is through cDNA synthesis, which has a very low success rate in generating a pure gene fragment. Therefore, it is of interest to devise a strategy to amplify a <i>P. falciparum</i> RPL12<sub>mito</sub> gene (PfRPL12<sub>mito</sub>) containing an intron through three sequential PCR steps using genomic DNA (gDNA) and primers with a minimum 15 bp 5'-overhang. The absence of an intronic segment and a continuous exonic region in the amplified gene fragment was confirmed using Sanger sequencing.
Authors
- Vikas Kumar (ORCID: https://orcid.org/0000-0002-6044-3239)
- Ankit Gupta (ORCID: https://orcid.org/0000-0002-0503-2322)
- Asad Abbas (ORCID: https://orcid.org/0000-0003-3752-058X)
- Manoj Kumar Yadav (ORCID: https://orcid.org/0000-0003-3219-0171)
Institutions
- All India Institute of Medical Sciences (IN)
Publication Details
- Journal
- Bioinformation
- Published
- 2025-08-31
- DOI
- https://doi.org/10.6026/973206300212278
- Primary Topic
- Molecular Biology Techniques and Applications
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- All-India Institute of Medical Sciences