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Related Articles

Sequencing:

Article Title: Two remarkable new species of Glaucocharis (Lepidoptera, Crambidae, Crambinae) from the Ogasawara Islands, Japan.
Article Snippet: we describe two new species of the genus Glaucocharis from the ogasawara Islands of Japan: G. triochellaris Matsui, Yagi & Hirowatari, sp. nov. and G. plumbofascialis Matsui, Yagi & Hirowatari, sp. nov. the new species with remarkable morphological characters are easily distinguished from congeneric species. we provide photographs of adult, male and female genitalia.. Molecular phylogenetic analysis revealed a sister relationship between the two new species and the monophyly of Glaucocharis.

Article Title: Redescription of immature stages of the shiitake fungus moth, Morophagoides moriutii Robinson (Lepidoptera: Tineidae)
Article Snippet: .. The amplified products were purified using ExoSAP–ITTM Express (Thermo Fisher Scientific Inc., USA), and sequencing was conducted at pre‐mixed Sanger sequencing services (Azenta, USA). ..

Article Title: Somatic EPAS1 Variants in Pheochromocytoma and Paraganglioma in Patients With Sickle Cell Disease.
Article Snippet: .. Purified products were sequenced in both forward and reverse directions using Sanger sequencing services provided by Azenta Life Sciences and sequences were analyzed using CLC Genomics Workbench 23 (QIAGEN, Aarhus, Denmark). ..

Article Title: A new Hibiscus-feeding species of Bucculatrix (Bucculatricidae) from the Ogasawara Islands, Japan
Article Snippet: The amplicons were purified using ExoSAPIT Express (Thermo Fisher Scientific Inc., Massachusetts, USA). .. The cycle sequences reactions and bidirectional sequencing were conducted using premixed Sanger sequencing services (Azenta Japan Corp., Tokyo, Japan). ..

Article Title: Improved protein binder design using β-pairing targeted RFdiffusion
Article Snippet: Protein binder designs were ordered as synthetic genes (eBlocks, Integrated DNA Technologies) and cloned via BsaI overhangs into the target cloning vector, LM0627 for Golden Gate assembly. .. Golden Gate reaction mixtures were transformed into a chemically competent expression strain (BL21(DE3)), and overnight outgrowth cultures were used to seed 500 mL protein expression cultures in auto-induction media (autoclaved TBII media supplemented with Kanamycin, 2 mM MgSO4, 1×5052) and to propagate expression plasmid for isolation (QIAprep Spin Miniprep Kit) and sanger sequencing services performed by GENEWIZ (Azenta Life Sciences). .. The following day (20-24 hrs later), cells were harvested and lysed by sonication (QSonica Q500 Sonicator), and clarified lysates were purified by immobilized metal affinity chromatography using Ni-NTA agarose resin (Qiagen).

Article Title: Genome reorganisation and expansion shape 3D genome architecture and define a distinct regulatory landscape in coleoid cephalopods
Article Snippet: PCR products were run on a 2% agarose (Sigma-Aldrich, A9793) gel and resulting bands were gel purified using the Monarch DNA Gel Extraction kit (NEB, T1020) following Manufacturer’s instructions. .. Purified PCR products were analyzed using Sanger Sequencing services provided by Genewiz (South Plainfield, NJ). ..

Article Title: Experimental evolution in the cystic fibrosis chemical environment reveals early TCA cycle flux as a central regulator of Mycobacterium abscessus biofilm formation
Article Snippet: .. PCR products with the correct fragment size were purified and sequenced using the Sanger Sequencing Services at Azenta Life Sciences. ..

Amplification:

Article Title: Redescription of immature stages of the shiitake fungus moth, Morophagoides moriutii Robinson (Lepidoptera: Tineidae)
Article Snippet: .. The amplified products were purified using ExoSAP–ITTM Express (Thermo Fisher Scientific Inc., USA), and sequencing was conducted at pre‐mixed Sanger sequencing services (Azenta, USA). ..

Purification:

Article Title: Redescription of immature stages of the shiitake fungus moth, Morophagoides moriutii Robinson (Lepidoptera: Tineidae)
Article Snippet: .. The amplified products were purified using ExoSAP–ITTM Express (Thermo Fisher Scientific Inc., USA), and sequencing was conducted at pre‐mixed Sanger sequencing services (Azenta, USA). ..

Article Title: Somatic EPAS1 Variants in Pheochromocytoma and Paraganglioma in Patients With Sickle Cell Disease.
Article Snippet: .. Purified products were sequenced in both forward and reverse directions using Sanger sequencing services provided by Azenta Life Sciences and sequences were analyzed using CLC Genomics Workbench 23 (QIAGEN, Aarhus, Denmark). ..

Article Title: Genome reorganisation and expansion shape 3D genome architecture and define a distinct regulatory landscape in coleoid cephalopods
Article Snippet: PCR products were run on a 2% agarose (Sigma-Aldrich, A9793) gel and resulting bands were gel purified using the Monarch DNA Gel Extraction kit (NEB, T1020) following Manufacturer’s instructions. .. Purified PCR products were analyzed using Sanger Sequencing services provided by Genewiz (South Plainfield, NJ). ..

Article Title: Experimental evolution in the cystic fibrosis chemical environment reveals early TCA cycle flux as a central regulator of Mycobacterium abscessus biofilm formation
Article Snippet: .. PCR products with the correct fragment size were purified and sequenced using the Sanger Sequencing Services at Azenta Life Sciences. ..

Transformation Assay:

Article Title: Improved protein binder design using β-pairing targeted RFdiffusion
Article Snippet: Protein binder designs were ordered as synthetic genes (eBlocks, Integrated DNA Technologies) and cloned via BsaI overhangs into the target cloning vector, LM0627 for Golden Gate assembly. .. Golden Gate reaction mixtures were transformed into a chemically competent expression strain (BL21(DE3)), and overnight outgrowth cultures were used to seed 500 mL protein expression cultures in auto-induction media (autoclaved TBII media supplemented with Kanamycin, 2 mM MgSO4, 1×5052) and to propagate expression plasmid for isolation (QIAprep Spin Miniprep Kit) and sanger sequencing services performed by GENEWIZ (Azenta Life Sciences). .. The following day (20-24 hrs later), cells were harvested and lysed by sonication (QSonica Q500 Sonicator), and clarified lysates were purified by immobilized metal affinity chromatography using Ni-NTA agarose resin (Qiagen).

Expressing:

Article Title: Improved protein binder design using β-pairing targeted RFdiffusion
Article Snippet: Protein binder designs were ordered as synthetic genes (eBlocks, Integrated DNA Technologies) and cloned via BsaI overhangs into the target cloning vector, LM0627 for Golden Gate assembly. .. Golden Gate reaction mixtures were transformed into a chemically competent expression strain (BL21(DE3)), and overnight outgrowth cultures were used to seed 500 mL protein expression cultures in auto-induction media (autoclaved TBII media supplemented with Kanamycin, 2 mM MgSO4, 1×5052) and to propagate expression plasmid for isolation (QIAprep Spin Miniprep Kit) and sanger sequencing services performed by GENEWIZ (Azenta Life Sciences). .. The following day (20-24 hrs later), cells were harvested and lysed by sonication (QSonica Q500 Sonicator), and clarified lysates were purified by immobilized metal affinity chromatography using Ni-NTA agarose resin (Qiagen).

Plasmid Preparation:

Article Title: Improved protein binder design using β-pairing targeted RFdiffusion
Article Snippet: Protein binder designs were ordered as synthetic genes (eBlocks, Integrated DNA Technologies) and cloned via BsaI overhangs into the target cloning vector, LM0627 for Golden Gate assembly. .. Golden Gate reaction mixtures were transformed into a chemically competent expression strain (BL21(DE3)), and overnight outgrowth cultures were used to seed 500 mL protein expression cultures in auto-induction media (autoclaved TBII media supplemented with Kanamycin, 2 mM MgSO4, 1×5052) and to propagate expression plasmid for isolation (QIAprep Spin Miniprep Kit) and sanger sequencing services performed by GENEWIZ (Azenta Life Sciences). .. The following day (20-24 hrs later), cells were harvested and lysed by sonication (QSonica Q500 Sonicator), and clarified lysates were purified by immobilized metal affinity chromatography using Ni-NTA agarose resin (Qiagen).

Isolation:

Article Title: Improved protein binder design using β-pairing targeted RFdiffusion
Article Snippet: Protein binder designs were ordered as synthetic genes (eBlocks, Integrated DNA Technologies) and cloned via BsaI overhangs into the target cloning vector, LM0627 for Golden Gate assembly. .. Golden Gate reaction mixtures were transformed into a chemically competent expression strain (BL21(DE3)), and overnight outgrowth cultures were used to seed 500 mL protein expression cultures in auto-induction media (autoclaved TBII media supplemented with Kanamycin, 2 mM MgSO4, 1×5052) and to propagate expression plasmid for isolation (QIAprep Spin Miniprep Kit) and sanger sequencing services performed by GENEWIZ (Azenta Life Sciences). .. The following day (20-24 hrs later), cells were harvested and lysed by sonication (QSonica Q500 Sonicator), and clarified lysates were purified by immobilized metal affinity chromatography using Ni-NTA agarose resin (Qiagen).

Polymerase Chain Reaction:

Article Title: Genome reorganisation and expansion shape 3D genome architecture and define a distinct regulatory landscape in coleoid cephalopods
Article Snippet: PCR products were run on a 2% agarose (Sigma-Aldrich, A9793) gel and resulting bands were gel purified using the Monarch DNA Gel Extraction kit (NEB, T1020) following Manufacturer’s instructions. .. Purified PCR products were analyzed using Sanger Sequencing services provided by Genewiz (South Plainfield, NJ). ..

Article Title: Experimental evolution in the cystic fibrosis chemical environment reveals early TCA cycle flux as a central regulator of Mycobacterium abscessus biofilm formation
Article Snippet: .. PCR products with the correct fragment size were purified and sequenced using the Sanger Sequencing Services at Azenta Life Sciences. ..

Mutagenesis:

Article Title: Optimization of the hongqu starter preparation process for the manufacturing of red mold rice with high gamma-aminobutyric acid production by solid-state fermentation.
Article Snippet: Anqi Wu and Liangyi Li contributed equally to this work.. Abstract Red mold rice (RMR) generally contains gamma-aminobutyric acid (GABA), which has several physiological functions.. Monascus purpureus M162, with a high GABA production of 15.10 mg/g was generated by atmospheric and room temperature plasma mutation.



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A minimal C-terminal helix in the RPGR BD is sufficient for CID binding and intercalates into the CID helical bundle through a conserved aromatic network. (A) Domain organization of human TTLL5 and RPGR. TTL catalytic core, blue; cMTBD, maroon; CID, green. RCC1-like β-propeller domain in RPGR, yellow, with each repeat shown as a separate propeller blade. <t>Sequence</t> for the minimal BD peptide required for TTLL5 targeting to RPGR shown in magenta at the top. (B) ITC measurements of WT miniBD peptide titrated into CID. Best fit of n = 2 independent experimental replicates. (C) 2.8-Å X-ray crystal structure of human TTLL5 CID in complex with human RPGR miniBD shown in cartoon representation; CID, green; RPGR, magenta. (D and E) Key conserved hydrophobic interactions at the CID–BD interface. Solid black boxes around residue labels designate disease-associated mutations, dashed boxes designate mutations without disease annotation in the ClinVar database that we predict are pathogenic based on our structure. RPGR and TTLL5 are colored as in B; H-bonds are shown as dashed lines.
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome <t>sequence</t> labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Image Search Results


A minimal C-terminal helix in the RPGR BD is sufficient for CID binding and intercalates into the CID helical bundle through a conserved aromatic network. (A) Domain organization of human TTLL5 and RPGR. TTL catalytic core, blue; cMTBD, maroon; CID, green. RCC1-like β-propeller domain in RPGR, yellow, with each repeat shown as a separate propeller blade. Sequence for the minimal BD peptide required for TTLL5 targeting to RPGR shown in magenta at the top. (B) ITC measurements of WT miniBD peptide titrated into CID. Best fit of n = 2 independent experimental replicates. (C) 2.8-Å X-ray crystal structure of human TTLL5 CID in complex with human RPGR miniBD shown in cartoon representation; CID, green; RPGR, magenta. (D and E) Key conserved hydrophobic interactions at the CID–BD interface. Solid black boxes around residue labels designate disease-associated mutations, dashed boxes designate mutations without disease annotation in the ClinVar database that we predict are pathogenic based on our structure. RPGR and TTLL5 are colored as in B; H-bonds are shown as dashed lines.

Journal: The Journal of Cell Biology

Article Title: Insights into retinal disease and non-tubulin glutamylation from a RPGR–TTLL5 complex structure

doi: 10.1083/jcb.202508020

Figure Lengend Snippet: A minimal C-terminal helix in the RPGR BD is sufficient for CID binding and intercalates into the CID helical bundle through a conserved aromatic network. (A) Domain organization of human TTLL5 and RPGR. TTL catalytic core, blue; cMTBD, maroon; CID, green. RCC1-like β-propeller domain in RPGR, yellow, with each repeat shown as a separate propeller blade. Sequence for the minimal BD peptide required for TTLL5 targeting to RPGR shown in magenta at the top. (B) ITC measurements of WT miniBD peptide titrated into CID. Best fit of n = 2 independent experimental replicates. (C) 2.8-Å X-ray crystal structure of human TTLL5 CID in complex with human RPGR miniBD shown in cartoon representation; CID, green; RPGR, magenta. (D and E) Key conserved hydrophobic interactions at the CID–BD interface. Solid black boxes around residue labels designate disease-associated mutations, dashed boxes designate mutations without disease annotation in the ClinVar database that we predict are pathogenic based on our structure. RPGR and TTLL5 are colored as in B; H-bonds are shown as dashed lines.

Article Snippet: Vectors were sequenced by Psomagen standard Sanger sequencing service.

Techniques: Binding Assay, Sequencing, Residue

Multiple sequence alignment of vertebrate TTLL5 CID and RPGR BD. (A) TTLL5 CID. (B) RPGR miniBD. Secondary structure elements indicated above the sequence and based on our X-ray crystal structure of the complex; gaps of disordered, unmodeled residues are indicated by a dotted line. Hydrophobic residues at the CID–BD interface indicated with blue boxes. Residues mutated for ITC experiments indicated with black arrows above the sequence.

Journal: The Journal of Cell Biology

Article Title: Insights into retinal disease and non-tubulin glutamylation from a RPGR–TTLL5 complex structure

doi: 10.1083/jcb.202508020

Figure Lengend Snippet: Multiple sequence alignment of vertebrate TTLL5 CID and RPGR BD. (A) TTLL5 CID. (B) RPGR miniBD. Secondary structure elements indicated above the sequence and based on our X-ray crystal structure of the complex; gaps of disordered, unmodeled residues are indicated by a dotted line. Hydrophobic residues at the CID–BD interface indicated with blue boxes. Residues mutated for ITC experiments indicated with black arrows above the sequence.

Article Snippet: Vectors were sequenced by Psomagen standard Sanger sequencing service.

Techniques: Sequencing

Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome sequence labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Biofilm

Article Title: Experimental evolution in the cystic fibrosis chemical environment reveals early TCA cycle flux as a central regulator of Mycobacterium abscessus biofilm formation

doi: 10.1016/j.bioflm.2025.100343

Figure Lengend Snippet: Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome sequence labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: PCR products with the correct fragment size were purified and sequenced using the Sanger Sequencing Services at Azenta Life Sciences.

Techniques: Variant Assay, Comparison, Sequencing, Binding Assay, Isolation