bl21 (New England Biolabs)
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BL21 Competent E coli
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BL21 Competent E coli 20x0 05 ml
Catalog Number:
c2530h
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204
Size:
1 ml
Category:
Competent Bacteria
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New England Biolabs
bl21
BL21 Competent E coli 20x0 05 ml
https://www.bioz.com/result/bl21/product/New England Biolabs
Average 95 stars, based on 81 article reviews
Price from $9.99 to $1999.99
BL21 Competent E coli 20x0 05 ml
https://www.bioz.com/result/bl21/product/New England Biolabs
Average 95 stars, based on 81 article reviews
Price from $9.99 to $1999.99
bl21 - by Bioz Stars,
2020-02
95/100 stars
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Clone Assay:Article Title: Algorithmic co-optimization of genetic constructs and growth conditions: application to 6-ACA, a potential nylon-6 precursor Article Snippet: Strains, plasmids and media Escherichia coli (E. coli ) DH5α was used for routine cloning and plasmid propagation (NEB, #C2987I). .. Article Title: RAB21 Activity Assay Using GST-fused APPL1 Article Snippet: .. 50 ml conical tube (Sarstedt AG & CO, catalog number: 62.547.004) 100 mm Tissue Culture Dish (Corning, catalog number: 430167) 0.22 μm filtering unit (Genesee Scientific Corporation, catalog number: 25-227) Amplification:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: G-blocks were PCR amplified using Forward primer 5′-TTATTCGAATTCGCGGCCGCTTCTAGAG and Reverse primer 5′-GGATTTCTGCAGCGGCCGCTACTAGTA with the following conditions: in a 50 μL reaction: 10 μL 5x HF buffer (New England Biolabs, NEB #E0553L), 2 μL 10 mM dNTP mix (NEB #N0447L), 1 μL 100 μM forward and reverse primer, 25 ng g block template, and 0.5 μL phusion polymerase. .. The ligation was transformed into chemically Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: The coding sequence of CsLOB1 of 714 bp was amplified with primers containing Eco RI/ Sal I restriction sites, and subcloned into the protein expression vector pGEX‐4T‐1, named pGEX‐4T‐1‐CsLOB1, using Q5® High‐Fidelity 2× Master Mix (New England Biolabs, Ipswich, MA, USA). .. The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Constructs containing the complete RNF114 sequence were amplified using primers containing 20 base pair homology to a pET24a plasmid (Novagen) that also contained a His8 -MBP-TEV sequence between Nde1 and BamH1 restriction sites. .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Filtration:Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Stable Transfection:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: Netrin-1 protein was either obtained from R & D (Minneapolis, MN, USA) or purified with anti-Myc tag affinity matrix from the conditioned media of HEK cells stably secreting netrin-1. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Article Title: RAB21 Activity Assay Using GST-fused APPL1 Article Snippet: .. 50 ml conical tube (Sarstedt AG & CO, catalog number: 62.547.004) 100 mm Tissue Culture Dish (Corning, catalog number: 430167) 0.22 μm filtering unit (Genesee Scientific Corporation, catalog number: 25-227) Synthesized:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: Gene cassettes were designed in CLC Main (Qiagen) and synthesized as g-blocks by IDT. .. The ligation was transformed into chemically Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: For the second method, DNA encoding the complete human isoform of RNF114 (Uniprot id: Q9Y508) was codon optimized for expression in E. coli and synthesized by Integrated DNA Technologies. .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Construct:Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin Article Snippet: GST Pull-Down Assays The GST-SMAD4 fusion protein was induced with 0.1 mM IPTG in Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: We have previously shown that these two katanin constructs have similar high severing activity (Bailey et al., ). .. We transformed the plasmid into Article Title: Proteome-wide enrichment of proteins modified by lysine methylation Article Snippet: Plasmids: 3×MBT-pGEX6P-1 and 3×MBTD355N -pGEX6P-1 constructs (AddGene plasmid nos. .. 46987 and 46988, respectively) Mammalian SILAC medium: DMEM (Thermo Scientific/Pierce, cat. no. 88420) and RPMI (Thermo Scientific/Pierce, cat. no. 88421) Dialyzed FBS for SILAC (Thermo Scientific/Pierce, cat. no. 88440) Yeast SILAC medium (based on Kubota et al. ): Dropout mix minus leucine, lysine, arginine and adenine (US Biological, cat. no. D9515F) Yeast nitrogen base without amino acids (Sigma, cat. no. Y0626) Leucine (Sigma, cat. no. L8912) Adenine (Sigma, cat. no. A8626) Glucose (Invitrogen-Gibco, cat. no. 15023021) SILAC amino acids: L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88429), L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 88427), 2 H4 -L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88438), 13 C6 -L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 88433), 15 N2 13 C6 -L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88209), 15 N4 13 C6 -L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 89990), L-proline (Thermo Scientific/Pierce, cat. no. 88430) Yeast strain from any background (for example, Fisher cat. no. NC0537738), but they must be auxotrophic for arginine and lysine ( arg4 lys2 ) as described in ref. LB medium (Sigma, cat. no. L3522) Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: We will refer to the Xenopus construct as Xl-p60 and the human construct as GFP-Hu-p60. .. The plasmid was transformed into Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: For TUBB3 RNAi rescue experiments, an RNAi-resistant construct was created by introducing seven silent point mutations in the target sequences. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Article Title: RAB21 Activity Assay Using GST-fused APPL1 Article Snippet: .. 50 ml conical tube (Sarstedt AG & CO, catalog number: 62.547.004) 100 mm Tissue Culture Dish (Corning, catalog number: 430167) 0.22 μm filtering unit (Genesee Scientific Corporation, catalog number: 25-227) Incubation:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Double-stranded (ds) RNAs for intererence in S2 cells have been generated using the following primers: T3-Snap29 5′-TAATACGACTCACTATAGGGAGA AACCCAGGAGGTGGGTAAG-3′ T7- Snap29 5′-AATTAACCCTCACTAAAGGGAGA ATGTTATCCAGCAATTCATTTTG-3′ DsRNA were in vitro transcribed with the T3 (Promega, P208C) and T7 (Promega, P207B) polymerase according to manufacturer's instructions, annealed, and incubated with the cells at a final concentration of 15ug/106 cells for 72 h. Multiple sequence analysis was performed using ClustalX using standard parameters. .. Antibody production GST-Snap29 expression was carried out in the Article Title: MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin Article Snippet: GST Pull-Down Assays The GST-SMAD4 fusion protein was induced with 0.1 mM IPTG in Activity Assay:Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: We have previously shown that these two katanin constructs have similar high severing activity (Bailey et al., ). .. We transformed the plasmid into Expressing:Article Title: Multivalent Antiviral XTEN–Peptide Conjugates with Long in Vivo Half-Life and Enhanced Solubility Article Snippet: .. XTEN Expression All of the thiol-containing XTEN precursors (XTEN-1, 2, 3, 4) were expressed in the Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: .. The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Paragraph title: Expression and Purification of Wild-type and C8A Mutant RNF114 Protein ... Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: .. Antibody production GST-Snap29 expression was carried out in the Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: Paragraph title: Generation and expression of GST fusion proteins ... Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: An isopropyl β -D-1 thiogalactopyranoside-inducible expression system was used for expression and purification. .. The plasmid was transformed into Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: The control was made by sham-purification from the conditioned media from HEK cells that had not been transfected with a cDNA expressing the Myc-tagged netrin-1. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Modification:Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: .. Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Transformation Assay:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: .. The ligation was transformed into chemically Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: .. The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin Article Snippet: .. GST Pull-Down Assays The GST-SMAD4 fusion protein was induced with 0.1 mM IPTG in Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: .. We transformed the plasmid into Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: .. Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: .. The plasmid was transformed into Article Title: Engineering transkingdom signalling in plants to control gene expression in rhizosphere bacteria Article Snippet: .. Protein purification For purification of IolG and IdhA, pOPS0141 and pOPS0142 were transformed into Transfection:Article Title: MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin Article Snippet: GST Pull-Down Assays The GST-SMAD4 fusion protein was induced with 0.1 mM IPTG in Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: The control was made by sham-purification from the conditioned media from HEK cells that had not been transfected with a cDNA expressing the Myc-tagged netrin-1. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Ligation:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: .. The ligation was transformed into chemically Protease Inhibitor:Article Title: RAB21 Activity Assay Using GST-fused APPL1 Article Snippet: .. 50 ml conical tube (Sarstedt AG & CO, catalog number: 62.547.004) 100 mm Tissue Culture Dish (Corning, catalog number: 430167) 0.22 μm filtering unit (Genesee Scientific Corporation, catalog number: 25-227) Cell Culture:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Generated:Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Double-stranded (ds) RNAs for intererence in S2 cells have been generated using the following primers: T3-Snap29 5′-TAATACGACTCACTATAGGGAGA AACCCAGGAGGTGGGTAAG-3′ T7- Snap29 5′-AATTAACCCTCACTAAAGGGAGA ATGTTATCCAGCAATTCATTTTG-3′ DsRNA were in vitro transcribed with the T3 (Promega, P208C) and T7 (Promega, P207B) polymerase according to manufacturer's instructions, annealed, and incubated with the cells at a final concentration of 15ug/106 cells for 72 h. Multiple sequence analysis was performed using ClustalX using standard parameters. .. Antibody production GST-Snap29 expression was carried out in the Droplet Countercurrent Chromatography:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Sequencing:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: .. The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Double-stranded (ds) RNAs for intererence in S2 cells have been generated using the following primers: T3-Snap29 5′-TAATACGACTCACTATAGGGAGA AACCCAGGAGGTGGGTAAG-3′ T7- Snap29 5′-AATTAACCCTCACTAAAGGGAGA ATGTTATCCAGCAATTCATTTTG-3′ DsRNA were in vitro transcribed with the T3 (Promega, P208C) and T7 (Promega, P207B) polymerase according to manufacturer's instructions, annealed, and incubated with the cells at a final concentration of 15ug/106 cells for 72 h. Multiple sequence analysis was performed using ClustalX using standard parameters. .. Antibody production GST-Snap29 expression was carried out in the Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: Plasmids were purified via MiniPrep (QIAGEN, Valencia, CA) before sequencing. .. Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: The target sequence was inserted into pAVU6+27 between Sal I site and Xba I site. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: .. Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Sonication:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: MyD88 Regulates the Expression of SMAD4 and the Iron Regulatory Hormone Hepcidin Article Snippet: .. GST Pull-Down Assays The GST-SMAD4 fusion protein was induced with 0.1 mM IPTG in Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: We transformed the plasmid into Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: The plasmid was transformed into Affinity Purification:Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Antibody production GST-Snap29 expression was carried out in the Binding Assay:Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: For data comparing depolymerization rates as a function of the concentration of katanin and quantifying katanin binding, we used a human p60 isoform with a green fluorescent protein (GFP) tag and maltose binding protein (MBP) tag for purification. .. We transformed the plasmid into Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: We also made an optimized human p60 construct with a maltose binding protein and GFP (GeneWiz, Cambridge, MA). .. The plasmid was transformed into Mutagenesis:Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Paragraph title: Expression and Purification of Wild-type and C8A Mutant RNF114 Protein ... Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: Point mutations (L113A, P116C, P116G, P116N, P117K, and P116S) and triple mutants (L113A/P116N/P117K, L113A/P116C/P117G, L113A/P116A/P117A) were generated using the Quick Change II XL Site-Directed Mutagenesis kit (Agilent Technologies, (Santa Clara, CA). .. Labeling:Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Purification:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: Purified, digested insert and vector were ligated with the following conditions 1 μL 10X T4 DNA ligase buffer (NEB), 50 ng vector, 150 ng insert, 0.5 μL T4 DNA ligase (NEB) in 10 μL total for 10 min at room temperature. .. The ligation was transformed into chemically Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Paragraph title: Expression and Purification of Wild-type and C8A Mutant RNF114 Protein ... Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: .. Antibody production GST-Snap29 expression was carried out in the Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: For data comparing depolymerization rates as a function of the concentration of katanin and quantifying katanin binding, we used a human p60 isoform with a green fluorescent protein (GFP) tag and maltose binding protein (MBP) tag for purification. .. We transformed the plasmid into Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: Plasmids were purified via MiniPrep (QIAGEN, Valencia, CA) before sequencing. .. Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: An isopropyl β -D-1 thiogalactopyranoside-inducible expression system was used for expression and purification. .. The plasmid was transformed into Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Article Title: Engineering transkingdom signalling in plants to control gene expression in rhizosphere bacteria Article Snippet: .. Protein purification For purification of IolG and IdhA, pOPS0141 and pOPS0142 were transformed into Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Protein Purification:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: Paragraph title: Protein purification ... The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: Paragraph title: Protein purification ... We transformed the plasmid into Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: Paragraph title: Protein purification ... The plasmid was transformed into Article Title: Engineering transkingdom signalling in plants to control gene expression in rhizosphere bacteria Article Snippet: .. Protein purification For purification of IolG and IdhA, pOPS0141 and pOPS0142 were transformed into Polymerase Chain Reaction:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. The ligation was transformed into chemically Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Gibson Assembly (NEB Gibson Assembly 2× Master Mix) was used to assemble the purified PCR product into the linearized vector. .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: The PCR product was inserted using BamHI and XhoI into pGEX -GST (Addgene Vector Database, 27- 4597- 01). .. Antibody production GST-Snap29 expression was carried out in the Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: PCR products were introduced into the pGEX-4T1 template between 5′ EcoRI and 3′XhoI sites. .. Affinity Chromatography:Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in Blocking Assay:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: G-blocks were PCR amplified using Forward primer 5′-TTATTCGAATTCGCGGCCGCTTCTAGAG and Reverse primer 5′-GGATTTCTGCAGCGGCCGCTACTAGTA with the following conditions: in a 50 μL reaction: 10 μL 5x HF buffer (New England Biolabs, NEB #E0553L), 2 μL 10 mM dNTP mix (NEB #N0447L), 1 μL 100 μM forward and reverse primer, 25 ng g block template, and 0.5 μL phusion polymerase. .. The ligation was transformed into chemically Recombinant:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Gel Extraction:Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: Products for PCR were assessed using 1% agarose gels (Invitrogen), and a QIAquick Gel Extraction kit (Qiagen) was used to purify PCR products of the correct length. .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically SDS Page:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Plasmid Preparation:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: Paragraph title: Construction of Plasmid DNA ... The ligation was transformed into chemically Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: .. The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Harnessing the Anti-Cancer Natural Product Nimbolide for Targeted Protein Degradation Article Snippet: .. Kanamycin (Kan)-resistant colonies were grown in LB media, and a Miniprep (Qiagen) kit was used to isolate the plasmid before sequence verification with appropriate primers. pET24a His8 –MBP plasmid (100 ng) containing the desired RNF114 construct was transformed into chemically Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: The PCR product was inserted using BamHI and XhoI into pGEX -GST (Addgene Vector Database, 27- 4597- 01). .. Antibody production GST-Snap29 expression was carried out in the Article Title: Algorithmic co-optimization of genetic constructs and growth conditions: application to 6-ACA, a potential nylon-6 precursor Article Snippet: Strains, plasmids and media Escherichia coli (E. coli ) DH5α was used for routine cloning and plasmid propagation (NEB, #C2987I). .. Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: .. We transformed the plasmid into Article Title: Inhibition of pressure-activated cancer cell adhesion by FAK-derived peptides Article Snippet: Generation and expression of GST fusion proteins Bacterial expression vector pGEX-4T1 (GE Healthcare, Munich, Germany) was used as a template to generate mutated and truncated human FAK as GST (Glutathione S-transferase) fusion proteins. .. Article Title: Proteome-wide enrichment of proteins modified by lysine methylation Article Snippet: Plasmids: 3×MBT-pGEX6P-1 and 3×MBTD355N -pGEX6P-1 constructs (AddGene plasmid nos. .. 46987 and 46988, respectively) Mammalian SILAC medium: DMEM (Thermo Scientific/Pierce, cat. no. 88420) and RPMI (Thermo Scientific/Pierce, cat. no. 88421) Dialyzed FBS for SILAC (Thermo Scientific/Pierce, cat. no. 88440) Yeast SILAC medium (based on Kubota et al. ): Dropout mix minus leucine, lysine, arginine and adenine (US Biological, cat. no. D9515F) Yeast nitrogen base without amino acids (Sigma, cat. no. Y0626) Leucine (Sigma, cat. no. L8912) Adenine (Sigma, cat. no. A8626) Glucose (Invitrogen-Gibco, cat. no. 15023021) SILAC amino acids: L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88429), L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 88427), 2 H4 -L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88438), 13 C6 -L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 88433), 15 N2 13 C6 -L-lysine-2HCl (Thermo Scientific/Pierce, cat. no. 88209), 15 N4 13 C6 -L-arginine-HCl (Thermo Scientific/Pierce, cat. no. 89990), L-proline (Thermo Scientific/Pierce, cat. no. 88430) Yeast strain from any background (for example, Fisher cat. no. NC0537738), but they must be auxotrophic for arginine and lysine ( arg4 lys2 ) as described in ref. LB medium (Sigma, cat. no. L3522) Article Title: Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails Article Snippet: .. The plasmid was transformed into Article Title: RAB21 Activity Assay Using GST-fused APPL1 Article Snippet: .. 50 ml conical tube (Sarstedt AG & CO, catalog number: 62.547.004) 100 mm Tissue Culture Dish (Corning, catalog number: 430167) 0.22 μm filtering unit (Genesee Scientific Corporation, catalog number: 25-227) Article Title: Forces during cellular uptake of viruses and nanoparticles at the ventral side Article Snippet: .. Production of titin-based tension probes Tension probes based on the I27 domain of titin were expressed in shRNA:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: TUBB3 shRNA and control shRNA were gifts from David L. Turner , EB3-GFP constructs were from Niels Galjart and TUBB3-V5 were from Elizabeth C Engle. .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Agarose Gel Electrophoresis:Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli Article Snippet: Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT). .. The ligation was transformed into chemically In Vitro:Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Double-stranded (ds) RNAs for intererence in S2 cells have been generated using the following primers: T3-Snap29 5′-TAATACGACTCACTATAGGGAGA AACCCAGGAGGTGGGTAAG-3′ T7- Snap29 5′-AATTAACCCTCACTAAAGGGAGA ATGTTATCCAGCAATTCATTTTG-3′ DsRNA were in vitro transcribed with the T3 (Promega, P208C) and T7 (Promega, P207B) polymerase according to manufacturer's instructions, annealed, and incubated with the cells at a final concentration of 15ug/106 cells for 72 h. Multiple sequence analysis was performed using ClustalX using standard parameters. .. Antibody production GST-Snap29 expression was carried out in the Produced:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: .. Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from Concentration Assay:Article Title: Functional characterization of the citrus canker susceptibility gene CsLOB1 Article Snippet: The expression vector pGEX‐4T‐1‐CsLOB1 was confirmed by sequencing, and transformed into Article Title: Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila Article Snippet: Double-stranded (ds) RNAs for intererence in S2 cells have been generated using the following primers: T3-Snap29 5′-TAATACGACTCACTATAGGGAGA AACCCAGGAGGTGGGTAAG-3′ T7- Snap29 5′-AATTAACCCTCACTAAAGGGAGA ATGTTATCCAGCAATTCATTTTG-3′ DsRNA were in vitro transcribed with the T3 (Promega, P208C) and T7 (Promega, P207B) polymerase according to manufacturer's instructions, annealed, and incubated with the cells at a final concentration of 15ug/106 cells for 72 h. Multiple sequence analysis was performed using ClustalX using standard parameters. .. Antibody production GST-Snap29 expression was carried out in the Article Title: Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails. Katanin catalyzes microtubule depolymerization independently of tubulin C‐terminal tails Article Snippet: For data comparing depolymerization rates as a function of the concentration of katanin and quantifying katanin binding, we used a human p60 isoform with a green fluorescent protein (GFP) tag and maltose binding protein (MBP) tag for purification. .. We transformed the plasmid into Staining:Article Title: Direct binding of TUBB3 with DCC couples netrin-1 signaling to intracellular microtubule dynamics in axon outgrowth and guidance Article Snippet: Recombinant DCC intracellular domain tagged with maltose-binding protein (DCC-ICD-MBP) was produced from |
