Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence
DNA barcoding is a molecular technique frequently used to identify or confirm a species, which involves the steps of isolation, amplification via PCR, and sequencing analysis. However, the use of lysate derived from samples after soaked and heated in TE buffer is lacking reported for this applicatio...
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| Format: | Article |
| Language: | English |
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EDP Sciences
2025-01-01
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| Series: | BIO Web of Conferences |
| Online Access: | https://www.bio-conferences.org/articles/bioconf/pdf/2025/06/bioconf_10thiccc_10006.pdf |
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| author | Su’udi Mukhamad Bahrul Ulum Fuad Setyati Dwi |
| author_facet | Su’udi Mukhamad Bahrul Ulum Fuad Setyati Dwi |
| author_sort | Su’udi Mukhamad |
| collection | DOAJ |
| description | DNA barcoding is a molecular technique frequently used to identify or confirm a species, which involves the steps of isolation, amplification via PCR, and sequencing analysis. However, the use of lysate derived from samples after soaked and heated in TE buffer is lacking reported for this application. This study aims to provide an alternative method for PCR using lysate as template for species identification of Bulbophyllum lobbii using rbcL primers. The results show that the lysate (after heating and briefly spun) is worthy of use as a template in PCR amplification, able to produce a thick single band with appropriate amplicon size (±600 bp). Further sequencing analysis confirms that the resulting sequence is highly readable with a clear chromatogram. BLAST analysis shows high identity (100%) with Bulbophyllum lobbii (MT518983) from USA. In sum, direct PCR using lysate provides an alternative approach for rapid DNA barcoding of plant samples with promising results. |
| format | Article |
| id | doaj-art-32354218800942f89ba5d0ee3541bf22 |
| institution | Kabale University |
| issn | 2117-4458 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | EDP Sciences |
| record_format | Article |
| series | BIO Web of Conferences |
| spelling | doaj-art-32354218800942f89ba5d0ee3541bf222025-02-05T10:43:24ZengEDP SciencesBIO Web of Conferences2117-44582025-01-011551000610.1051/bioconf/202515510006bioconf_10thiccc_10006Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequenceSu’udi Mukhamad0Bahrul Ulum Fuad1Setyati Dwi2Biology Department, Faculty of Mathematics and Natural Sciences, University of JemberBiology Department, Faculty of Mathematics and Natural Sciences, University of JemberBiology Department, Faculty of Mathematics and Natural Sciences, University of JemberDNA barcoding is a molecular technique frequently used to identify or confirm a species, which involves the steps of isolation, amplification via PCR, and sequencing analysis. However, the use of lysate derived from samples after soaked and heated in TE buffer is lacking reported for this application. This study aims to provide an alternative method for PCR using lysate as template for species identification of Bulbophyllum lobbii using rbcL primers. The results show that the lysate (after heating and briefly spun) is worthy of use as a template in PCR amplification, able to produce a thick single band with appropriate amplicon size (±600 bp). Further sequencing analysis confirms that the resulting sequence is highly readable with a clear chromatogram. BLAST analysis shows high identity (100%) with Bulbophyllum lobbii (MT518983) from USA. In sum, direct PCR using lysate provides an alternative approach for rapid DNA barcoding of plant samples with promising results.https://www.bio-conferences.org/articles/bioconf/pdf/2025/06/bioconf_10thiccc_10006.pdf |
| spellingShingle | Su’udi Mukhamad Bahrul Ulum Fuad Setyati Dwi Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence BIO Web of Conferences |
| title | Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence |
| title_full | Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence |
| title_fullStr | Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence |
| title_full_unstemmed | Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence |
| title_short | Direct PCR for DNA barcoding of Bulbophyllum lobbii Lindl. based on rbcL sequence |
| title_sort | direct pcr for dna barcoding of bulbophyllum lobbii lindl based on rbcl sequence |
| url | https://www.bio-conferences.org/articles/bioconf/pdf/2025/06/bioconf_10thiccc_10006.pdf |
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