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TaKaRa stellar competent cells
Stellar Competent Cells, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/stellar competent cells/product/TaKaRa
Average 99 stars, based on 74 article reviews
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stellar competent cells - by Bioz Stars, 2020-04
99/100 stars

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Clone Assay:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Paragraph title: Construction of full-length infectious cDNA clones of PeWBVYV and transmission by whiteflies. ... A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Paragraph title: Amplification and Cloning. ... Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766). .. Positive clones were recovered and verified via Sanger sequencing.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA). .. To produce Sox2-t2A-mCherry, Sox2-t2A-GFP was then cut with KpnI and AgeI to remove GFP and replace it with PCR-amplified mCherry from pAAVS1-NDi-CRISPRi (Addgene, plasmid no. 73497; a gift from Bruce Conklin) ( ) with KpnI and AgeI overhang primers.

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: To generate the pooled double sgRNA library, the sgRNA sequence were PCR amplified from pooled pSLQ5004-sgRNA constructs, gel purified, and ligated onto the Nsil-digested pooled pSLQ1373-sgRNA constructs using In- Fusion cloning (Clontech). .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Amplification:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Briefly, the three fragments showing overlap were PCR amplified separately from cDNA constructed from an infected sample using Q5 High-fidelity DNA polymerase (NEB) and subjected to gel purification, and 120-ng volumes of the fragments were mixed. .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Paragraph title: Amplification and Cloning. ... Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: To generate the pooled double sgRNA library, the sgRNA sequence were PCR amplified from pooled pSLQ5004-sgRNA constructs, gel purified, and ligated onto the Nsil-digested pooled pSLQ1373-sgRNA constructs using In- Fusion cloning (Clontech). .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Synthesized:

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: We synthesized individual oligos encoding sgRNAs in a 96-well format (IDT), and cloned each sgRNA into pSLQ1373 individually as previously described. .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Construct:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Briefly, the three fragments showing overlap were PCR amplified separately from cDNA constructed from an infected sample using Q5 High-fidelity DNA polymerase (NEB) and subjected to gel purification, and 120-ng volumes of the fragments were mixed. .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: Next, dCas9-KRAB was excised out using NheI/XbaI to create a lineralized CMV-t2a-GFP construct. .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA).

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: To generate the pooled double sgRNA library, the sgRNA sequence were PCR amplified from pooled pSLQ5004-sgRNA constructs, gel purified, and ligated onto the Nsil-digested pooled pSLQ1373-sgRNA constructs using In- Fusion cloning (Clontech). .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Electrophoresis:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: The final PCR program was run as follows: 1 cycle of 98 °C for 10 s; 35 cycles of 98 °C for 10 s, 67 °C for 30 s, 72 °C for 1 min; 1 cycle of 72 °C for 5 min. PCR products were separated on 1.5% agarose gels by electrophoresis and visualized, using ethidium bromide staining. .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Incubation:

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: The In-Fusion reaction mixture was incubated at 37 °C for 15 min, followed by 50 °C for 15 min, and was then cooled on ice. .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Expressing:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Paragraph title: Construction of the recombinant expression vector ... Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Transformation Assay:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin. .. Colonies were screened for full-length virus by colony PCR using primers ( ) followed by plasmid extraction and sequencing from positive clones.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech). .. Of the 100-μL transformation mixture [after addition of 80 μL of SOC medium (Thermo Fisher Scientific)], 2 μL was diluted with 23 μL of SOC medium, spread on selective medium [LB agar + ampicillin (50 µg/mL)], and incubated overnight at 37 °C.

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight. .. At least 48 isolated Escherichia coli colonies were screened using colony PCR with primers that surround the site of integration in the pLEXSY vector (P1442 and A264).

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: .. PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766). .. Positive clones were recovered and verified via Sanger sequencing.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA). .. To produce Sox2-t2A-mCherry, Sox2-t2A-GFP was then cut with KpnI and AgeI to remove GFP and replace it with PCR-amplified mCherry from pAAVS1-NDi-CRISPRi (Addgene, plasmid no. 73497; a gift from Bruce Conklin) ( ) with KpnI and AgeI overhang primers.

Gel Purification:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Briefly, the three fragments showing overlap were PCR amplified separately from cDNA constructed from an infected sample using Q5 High-fidelity DNA polymerase (NEB) and subjected to gel purification, and 120-ng volumes of the fragments were mixed. .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Transfection:

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: Paragraph title: Plasmids, transfections, and enzymes. ... PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766).

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: Paragraph title: Transient transfections ... The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA).

Ligation:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin. .. Colonies were screened for full-length virus by colony PCR using primers ( ) followed by plasmid extraction and sequencing from positive clones.

Infection:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Briefly, the three fragments showing overlap were PCR amplified separately from cDNA constructed from an infected sample using Q5 High-fidelity DNA polymerase (NEB) and subjected to gel purification, and 120-ng volumes of the fragments were mixed. .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Generated:

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: UL138 mutants were generated in pSG5 from a WT clone ( ). .. PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766).

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: Sox2-t2A-GFP was generated by amplifying the human SOX2 sequence from a donor construct (HsCD00079917, Harvard Institute of Proteomics) ( ) using NheI and XbaI overhang primers and Phusion high-fidelity polymerase (New England Biolabs). .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA).

Transmission Assay:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: Paragraph title: Construction of full-length infectious cDNA clones of PeWBVYV and transmission by whiteflies. ... A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin.

Polymerase Chain Reaction:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin. .. Colonies were screened for full-length virus by colony PCR using primers ( ) followed by plasmid extraction and sequencing from positive clones.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Briefly, 10–50 ng of purified PCR product (2 μL postelution) was mixed with 50 ng of linearized pGL4.23 vector in a total volume of 10 μL, added to the In-Fusion HD EcoDry pellet, and mixed by pipetting. .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: PCR products were purified, using the NucleoSpin PCR Clean-Up Kit (Macherey-Nagel, GmbH & Co. KG, Germany), according to the manufacturer’s instructions. .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: .. PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766). .. Positive clones were recovered and verified via Sanger sequencing.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA). .. To produce Sox2-t2A-mCherry, Sox2-t2A-GFP was then cut with KpnI and AgeI to remove GFP and replace it with PCR-amplified mCherry from pAAVS1-NDi-CRISPRi (Addgene, plasmid no. 73497; a gift from Bruce Conklin) ( ) with KpnI and AgeI overhang primers.

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: To generate the pooled double sgRNA library, the sgRNA sequence were PCR amplified from pooled pSLQ5004-sgRNA constructs, gel purified, and ligated onto the Nsil-digested pooled pSLQ1373-sgRNA constructs using In- Fusion cloning (Clontech). .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Recombinant:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Paragraph title: Construction of the recombinant expression vector ... Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Nucleic Acid Electrophoresis:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight. .. Using gel electrophoresis, all colonies that contained inserts larger than 300 bp were selected for further subculture in 5 ml LB medium overnight.

Magnetic Beads:

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: PCR products were purified using AMPure XP magnetic beads (Beckman Coulter) following the manufacturer’s recommendations, and the final elution was performed using 30 μL of 10 mM Tris pH 8.0. .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Mutagenesis:

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: Briefly, the mutagenesis primers listed in were used to amplify UL138HA-pSG5 ( ) using CloneAmp HiFi polymerase (Clontech; 639298). .. PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766).

Isolation:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight. .. At least 48 isolated Escherichia coli colonies were screened using colony PCR with primers that surround the site of integration in the pLEXSY vector (P1442 and A264).

Purification:

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Briefly, 10–50 ng of purified PCR product (2 μL postelution) was mixed with 50 ng of linearized pGL4.23 vector in a total volume of 10 μL, added to the In-Fusion HD EcoDry pellet, and mixed by pipetting. .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Next, the purified amplicons were integrated into a Xba I or Nco I and Kpn I double digested pLEXSY-hyg2 vector, using In-Fusion recombinase. .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA). .. To produce Sox2-t2A-mCherry, Sox2-t2A-GFP was then cut with KpnI and AgeI to remove GFP and replace it with PCR-amplified mCherry from pAAVS1-NDi-CRISPRi (Addgene, plasmid no. 73497; a gift from Bruce Conklin) ( ) with KpnI and AgeI overhang primers.

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen). .. Primers (IDT) used to construct individual sgRNAs are shown in .

Sequencing:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin. .. Colonies were screened for full-length virus by colony PCR using primers ( ) followed by plasmid extraction and sequencing from positive clones.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech). .. Positive clones for each allele of each variant were identified by restriction digestion (KpnI and XhoI) and Sanger sequencing of plasmid DNA using pGL4.23 vector backbone primers.

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight. .. Plasmids were harvested using the QIAprep Spin Miniprep (Qiagen) and sent for bi-directional sequencing (VIB Genetic Sequencing Facility, Antwerp, Belgium) using the P1442 and A246 primers.

Article Title: The Membrane-Spanning Peptide and Acidic Cluster Dileucine Sorting Motif of UL138 Are Required To Downregulate MRP1 Drug Transporter Function in Human Cytomegalovirus-Infected Cells
Article Snippet: PCR products were digested with DpnI enzyme (NEB; R0176S) and transformed into Stellar competent cells (Clontech; 636766). .. Positive clones were recovered and verified via Sanger sequencing.

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: Sox2-t2A-GFP was generated by amplifying the human SOX2 sequence from a donor construct (HsCD00079917, Harvard Institute of Proteomics) ( ) using NheI and XbaI overhang primers and Phusion high-fidelity polymerase (New England Biolabs). .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA).

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: To generate the pooled double sgRNA library, the sgRNA sequence were PCR amplified from pooled pSLQ5004-sgRNA constructs, gel purified, and ligated onto the Nsil-digested pooled pSLQ1373-sgRNA constructs using In- Fusion cloning (Clontech). .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen).

Staining:

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: The final PCR program was run as follows: 1 cycle of 98 °C for 10 s; 35 cycles of 98 °C for 10 s, 67 °C for 30 s, 72 °C for 1 min; 1 cycle of 72 °C for 5 min. PCR products were separated on 1.5% agarose gels by electrophoresis and visualized, using ethidium bromide staining. .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight.

Plasmid Preparation:

Article Title: Transmission of a New Polerovirus Infecting Pepper by the Whitefly Bemisia tabaci
Article Snippet: .. A 240-ng volume of the gel-purified final PCR product was ligated to 120 ng of PCR-linearized ( ) pJL89 binary vector by the use of Gibson assembly mastermix (NEB) for 2 h. A 2-μl volume of the ligation mixture was transformed to Stellar competent cells (TaKaRa) and grown overnight on LB agar with kanamycin. .. Colonies were screened for full-length virus by colony PCR using primers ( ) followed by plasmid extraction and sequencing from positive clones.

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Briefly, 10–50 ng of purified PCR product (2 μL postelution) was mixed with 50 ng of linearized pGL4.23 vector in a total volume of 10 μL, added to the In-Fusion HD EcoDry pellet, and mixed by pipetting. .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Article Title: Expression of a rK39 homologue from an Iranian Leishmania infantum isolate in Leishmania tarentolae for serodiagnosis of visceral leishmaniasis
Article Snippet: .. Finally, the completed vector was transformed into Stellar™ Competent Cells (Takara Bio Company, CA, USA) by heat shock and plated on Luria Bertani (LB) agar supplemented with 100 µg/ml carbenicillin at 37 °C overnight. .. At least 48 isolated Escherichia coli colonies were screened using colony PCR with primers that surround the site of integration in the pLEXSY vector (P1442 and A264).

Article Title: KLF4 protein stability regulated by interaction with pluripotency transcription factors overrides transcriptional control
Article Snippet: A t2a-GFP backbone was subcloned from pLV hU6-sgRNA hUbC-dCas9-KRAB-t2a-GFP (Addgene plasmid no. 71237, a gift from Charles Gersbach) ( ) and inserted into hCas9 (Addgene, plasmid no. 41815; a gift from George Church) ( ) using AgeI/XbaI digestion and T4 DNA ligase (Thermo Fisher Scientific). .. The SOX2 PCR product was purified and inserted into CMV-t2a-GFP using the In-Fusion HD cloning kit (Takara Bio USA) and transformed into Stellar-Competent cells (Takara Bio USA).

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: .. The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen). .. Primers (IDT) used to construct individual sgRNAs are shown in .

Next-Generation Sequencing:

Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
Article Snippet: The double sgRNA-library pools were prepared in Stellar competent cells (TaKaRa) and purified with a Plasmid Maxi Kit (Qiagen). .. The representation of each of the double-sgRNA constructs was then quantified by NGS with the primers listed in .

Gel Extraction:

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: The pGL4.23 vector (Promega) was linearized by double digestion with KpnI-HF (New England Biolabs) and XhoI (New England Biolabs) and gel-purified using a QIAquick Gel Extraction Kit (Qiagen). .. Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech).

Variant Assay:

Article Title: INAUGURAL ARTICLE by a Recently Elected Academy Member:Multiple SCN5A variant enhancers modulate its cardiac gene expression and the QT interval
Article Snippet: Following a 10-fold dilution with 10 mM Tris pH 8.0, 1 μL of the diluted In-Fusion reaction mixture was used to transform 20 μL of Stellar competent cells (Clontech). .. Positive clones for each allele of each variant were identified by restriction digestion (KpnI and XhoI) and Sanger sequencing of plasmid DNA using pGL4.23 vector backbone primers.

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    TaKaRa stella competent escherichia coli cells
    Stella Competent Escherichia Coli Cells, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stella competent escherichia coli cells/product/TaKaRa
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    stella competent escherichia coli cells - by Bioz Stars, 2020-04
    99/100 stars
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