avai 10u  (New England Biolabs)

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Live-cell imaging reveals decreased cAMP in a PFE-associated c.1050-3C>G PTH1R cell model

doi: 10.1007/s00109-026-02668-8

Figure Lengend Snippet: Genomic analysis of the PDL-hTERT cell line with heterozygous c.1050-3C > G PTH1R variant. A The upper two sequences show the forward (fw) and reverse (rv) wild-type gDNA sequence, while the two lower sequences display the genetically modified forward and reverse gDNA sequence. The red arrow and the blue line indicate the c.1050-3 position showing a C in the wild-type sequence and a double peak, containing a C and a G in the genetically modified sequence. The guide RNA (gRNA) as well as the PAM sequence are highlighted on top of the sequences. B The upper part illustrates the AvaI restriction enzyme cutting side resulting from the base exchange in general. The lower part shows the newly created AvaI restriction enzyme cutting side in the PTH1R sequence as it is a heterozygous mutation with only one allele showing the WT sequence (blue box) and the other allele represents the mutated allele (red box). C Agarose gel with a 1-kb ladder in the first column, the WT PCR product digested without (−) and with (+) AvaI in the second and third columns and the digested genetically modified PCR product without (−) and with (+) AvaI in the fourth and fifth columns. The blue box indicates the 702 bp PCR products containing the wild type sequence, while the red box shows the one digested product of the PCR with a size of 528 bp and the green box displays the other digested product of the PCR with 174 bp which indicates the exchange from C to G

Article Snippet: The c.1050-3C>G substitution creates an AvaI restriction site, enabling mutation detection by digesting PCR products with AvaI (10 U/ml, New England Biolabs), and separating them on an agarose gel.

Techniques: Variant Assay, Sequencing, Genetically Modified, Mutagenesis, Agarose Gel Electrophoresis

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Live-cell imaging reveals decreased cAMP in a PFE-associated c.1050-3C>G PTH1R cell model

doi: 10.1007/s00109-026-02668-8

Figure Lengend Snippet: Genomic analysis of the PDL-hTERT cell line with heterozygous c.1050-3C > G PTH1R variant. A The upper two sequences show the forward (fw) and reverse (rv) wild-type gDNA sequence, while the two lower sequences display the genetically modified forward and reverse gDNA sequence. The red arrow and the blue line indicate the c.1050-3 position showing a C in the wild-type sequence and a double peak, containing a C and a G in the genetically modified sequence. The guide RNA (gRNA) as well as the PAM sequence are highlighted on top of the sequences. B The upper part illustrates the AvaI restriction enzyme cutting side resulting from the base exchange in general. The lower part shows the newly created AvaI restriction enzyme cutting side in the PTH1R sequence as it is a heterozygous mutation with only one allele showing the WT sequence (blue box) and the other allele represents the mutated allele (red box). C Agarose gel with a 1-kb ladder in the first column, the WT PCR product digested without (−) and with (+) AvaI in the second and third columns and the digested genetically modified PCR product without (−) and with (+) AvaI in the fourth and fifth columns. The blue box indicates the 702 bp PCR products containing the wild type sequence, while the red box shows the one digested product of the PCR with a size of 528 bp and the green box displays the other digested product of the PCR with 174 bp which indicates the exchange from C to G

Article Snippet: The c.1050-3C>G substitution creates an AvaI restriction site, enabling mutation detection by digesting PCR products with AvaI (10 U/ml, New England Biolabs), and separating them on an agarose gel.

Techniques: Variant Assay, Sequencing, Genetically Modified, Mutagenesis, Agarose Gel Electrophoresis