✦ LIBER ✦

Direct sequencing of tobacco chloroplast genome by the polymerase chain reaction

✍ Scribed by Kin-Ying To; Chiu-Yueh Li; Yu-Sun Chang; Shih-Tung Liu

Publisher: Springer
Year: 1992
Tongue: English
Weight: 599 KB
Volume: 19
Category: Article
ISSN: 0167-4412
DOI: 10.1007/bf00040540

No coin nor oath required. For personal study only.

✦ Synopsis

We have developed a polymerase chain reaction (PCR) method for sequencing of tobacco chloroplast genome. In a mixture containing chloroplast DNA, 5'-end-labeled oligonucleotide primer, Taq DNA polymerase and reaction buffer, we were able to sequence a segment of chloroplast 16S rRNA gene. The results showed that the 750 bp of DNA sequenced were identical to the sequence reported, indicating that direct sequencing method that we have developed is useful for the sequencing of chloroplast genome. To analyze the chloroplast genome more rapidly in those in vitro grown plantlets, we also developed a simple method which is applicable for the amplifications and sequencing of chloroplast 16S rRNA fragment from either 0.15 g of tobacco leaf or stem tissue. The readable sequences obtained from the presented methods were consistent with the published sequence.

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