Obtain the annotated sequence of mRFP-Rab5 from Addgenes. If you load the annotations (in Genbank format) SnapGene will automatically show them. Genbank format not only contains a sequence but also all annotations. Even if you don't load them from Genbank it is still a good idea to import them in Genbank format (and not in Fasta format). In SnapGene you can load sequences directly from Genbank or you can load them by importing a file. Fortunately, many vectors are available from Addgene, an organization that improves sharing of plasmids. However, not all vector sequences are stored in the SnapGene database. Loading vector sequences from other resources Go back to SnapGene and click the Open button in the top menu:īrowse to the pAcGFP1-C1.dna file and open it. On the webpage of pAcGFP1-C1 click pAcGFP1-C1.dna: Try to load it without peeking at the solution. This vector is available in SnapGene's list of fluorescent protein genes and plasmids. You now see the circular view of the vector with restriction sites automatically visualized:Įxercise: load the sequence of vector pAcGFP1-C1 Open SnapGene and select Open in the startup menu:īrowse to the pcDNA4 TO.dna file and open it. Open the annotated sequence of pcDNA4/TO in SnapGene. On the webpage of pcDNA4/TO click the pcDNA4 TO.dna link to save the file: Hence, awareness for uptake of cancer screening and early detection are encouraged.Annotation: every possible piece of information about a sequenceĭownload the annotated sequence of pcDNA4/TO from SnapGene. Our study also revealed the poor knowledge of Nigerian women to cancer screening, this may be one of the contributory factors to late diagnosis of cervical cancer among the African population. The histopathological result, alongside the Multiple Sequence Alignment revealed that with higher severity of cervical carcinoma, the p53 gene tends to accumulate more mutations. Sociodemographic characteristics revealed that 60% of the study participants has husbands with multiple sexual partners and that only 23.3% of the participants have ever done pap smear test prior to diagnosis, while 20% of them are unaware of the screening test. The outcome of this study showed nineteen identified polymorphic variants Missense mutations occurred in 47% of the samples, 32% were silent mutations, 16% were frameshift mutation and 5% nonsense mutation. Multiple Sequence Alignment using T-COFFEE was performed to reveal the polymorphic variations in the samples. Nucleotide BLAST was performed on the DNA sequence for sequence identity and retrieval of reference sequence.
The PCR products were sequenced at Inqaba Biotec, and chromatogram was analyzed using FinchTv and SnapGene Viewer. The region of interest was amplified using Polymerase Chain Reaction (PCR), this was quantified on a 1.5% agarose gel, and the ethidium bromide stained gel was viewed on a transilluminator. The purified genomic DNA was quantified using NanoDropTM 2000 spectrophotometer. Genomic DNA was isolated and purified from cervical tissue biopsies obtained from patients reporting at the University College Hospital, Ibadan. This research study explored the mutations in the exon 3 and exon 4 of the P53 gene in cervical cancer.
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