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Bio-Rad genepulser xcell
Genepulser Xcell, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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genepulser xcell - by Bioz Stars, 2020-01
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Related Articles

Clone Assay:

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: MRPA was amplified using iProof high-fidelity DNA polymerase (Bio-Rad) with GC buffer plus 10% dimethyl sulfoxide (DMSO) and directly cloned in the pLEXSY-hyg2 using the In-Fusion HD cloning kit (Clontech). .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ).

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Both the 5’-flanking and 3’-flanking PCR products were cloned upstream and downstream of the hygromycin (HYG) or neomycin (NEO) drug cassette respectively, in pX-63-HYG and pX-63-NEO vectors. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: Paragraph title: Cloning, expression and purification of recombinant VTN-1 to -4 and -2-1 to -2-3 ... All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

Centrifugation:

Article Title: RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene
Article Snippet: Trophozoites were grown at 37 °C in TYI-S33 culture medium; the cells were harvested at mid-to-late logarithmic phase by chilling the culture tubes on ice for 10 min and collected by centrifugation (1000× g /10 min at 4 °C). .. Upon placing a concentrated cell suspension (300 μL containing 1 × 107 cells) in a previously chilled 0.4 cm-gap electroporation cuvette, plasmidic DNA (50 μg) was added, and the mixture was incubated on ice for 5 min. Next, cells were electroporated using the following conditions: 350 V, 1000 μF and 700 Ω in a GenePulser Xcell (Bio-rad, Hercules, CA, USA).

Amplification:

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: MRPA was amplified using iProof high-fidelity DNA polymerase (Bio-Rad) with GC buffer plus 10% dimethyl sulfoxide (DMSO) and directly cloned in the pLEXSY-hyg2 using the In-Fusion HD cloning kit (Clontech). .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ).

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Similarly 3’-flanking region (775 bp) was PCR amplified using F8/R8 primer pair ( ) containing a Sma I or Bgl II restriction endonuclease site. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: The amplified fragments were digested and ligated in-frame into the Eco RI and Not I sites of the pPICZα A expression vector for Pichia pastoris Expression System (Thermo Fisher Scientific). .. All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

Mass Spectrometry:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: .. Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA). .. Next, cells were suspended in 400 µl of Ingenio™ Electroporation Solution (Mirus Bio LLC, MC-MIR-50114) with 4 nmol (for 4 wells) of Atg7 or NS siRNA and electroporation was performed in accordance with the manufacturer's instructions using the optimal program (170 V, 10 ms, 2 pulses).

Article Title: On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei
Article Snippet: .. Then, 25 μg of linearized plasmid DNA was added to the solution and cells were pulsed twice at 1,600 V with a time constant of 0.6 ms on a GenePulser Xcell (BioRad, Hercules, CA, USA). ..

Stable Transfection:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Construct:

Article Title: Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization
Article Snippet: HA-tagged DAPs for integrated expression in Giardia were constructed by amplifying 86676 (δ-Giardin) or 16263 from WBC6 genomic DNA and inserted into pKS_3HA_Pur (linearized by NotI and AflII) by Gibson Assembly. .. After construction, 86676_3HA_Pur or 16263_3HA_Pur were linearized for homologous recombination by PstI or NsiI, respectively and ~10 µg of plasmid was electroporated into 300 µl of Giardia in a 0.4 cm cuvette using a GenePulser XCell (BioRad) set to 375 V, 1000 µF and 750 ohms.

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: The plasmid p-VBIG was constructed by inserting a 2A- IDI -2A- GPS DNA fragment into the pUC18-VHb-ble vector [ ] by Gibson Assembly Master Mix (NEB, Ipswich, MA, USA). .. The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA).

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: .. All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA). .. Recombinants were expressed and purified by His GraviTrap (GE Healthcare) according to the manufacturer's protocol.

Incubation:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: For the former, plasmid DNA was added to TurboFectin 8.0 in OptiMEM I (TurboFectin:DNA, 3:1), incubated (15-30 min, 20°C), and the complex was then added to cells in 6-well plates (1-1.5 μg DNA/well) in complete medium, and incubated for 24-48 h. For electroporation, cells (80%–90% confluent in a T75 flask) were scraped into culture medium, centrifuged (150 xg , 5 min), and resuspended in Opti-MEM I (2 × 106 cells/mL). .. Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms).

Article Title: RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene
Article Snippet: .. Upon placing a concentrated cell suspension (300 μL containing 1 × 107 cells) in a previously chilled 0.4 cm-gap electroporation cuvette, plasmidic DNA (50 μg) was added, and the mixture was incubated on ice for 5 min. Next, cells were electroporated using the following conditions: 350 V, 1000 μF and 700 Ω in a GenePulser Xcell (Bio-rad, Hercules, CA, USA). ..

Expressing:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Article Title: Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization
Article Snippet: HA-tagged DAPs for integrated expression in Giardia were constructed by amplifying 86676 (δ-Giardin) or 16263 from WBC6 genomic DNA and inserted into pKS_3HA_Pur (linearized by NotI and AflII) by Gibson Assembly. .. After construction, 86676_3HA_Pur or 16263_3HA_Pur were linearized for homologous recombination by PstI or NsiI, respectively and ~10 µg of plasmid was electroporated into 300 µl of Giardia in a 0.4 cm cuvette using a GenePulser XCell (BioRad) set to 375 V, 1000 µF and 750 ohms.

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: Paragraph title: Cloning, expression and purification of recombinant VTN-1 to -4 and -2-1 to -2-3 ... All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

Transformation Assay:

Article Title: Construction of a Bile-responsive Expression System inLactobacillus plantarum
Article Snippet: Paragraph title: DNA isolation, transformation, and bile induction ... This mixture was subjected to electroporation using GenePulser Xcell™ (BioRad, USA) using 200 Ω, 2.0 kV and 25 μF capacitance conditions.

Article Title: A non-catalytic role of RecBCD in homology directed gap repair and translesion synthesis
Article Snippet: Monitoring HDGR and TLS To 40 μl aliquot of competent cells, prepared as previously described , 1 ng of the lesion-carrying vector mixed with 1 ng of the internal standard (pVP146) was added and electroporated in a GenePulser Xcell from BioRad (2.5 kV, 25 μF, 200 Ω). .. Part of the cells were plated on LB + 10 μg/ml tetracycline to measure the transformation efficiency of plasmid pVP146, and the rest were plated on LB + 50 μg/ml ampicillin + 80 μg/ml X-gal to select for integrants (AmpR ) and to visualize HDGR or TLS events (lacZ+ phenotype depending on the vector used).

Article Title: Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization
Article Snippet: Paragraph title: Vector Construction and Giardia Transformation ... After construction, 86676_3HA_Pur or 16263_3HA_Pur were linearized for homologous recombination by PstI or NsiI, respectively and ~10 µg of plasmid was electroporated into 300 µl of Giardia in a 0.4 cm cuvette using a GenePulser XCell (BioRad) set to 375 V, 1000 µF and 750 ohms.

Article Title: Consequences of Cas9 cleavage in the chromosome of Escherichia coli
Article Snippet: .. The pCas9, pCas9::LacZ1 and pCas9::LacZ2 plasmids (40 ng each) were transformed into electrocompetent cells using 1 mm gap cuvette and the GenePulser Xcell (Bio-Rad) set to 1800 V, 25 uF, 200 Ω. ..

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: .. The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA). .. The parameters of electroporation were set as 2 kV, 50 μF, and 300 Ω. Electroporated cells were resuspended in 50 mL of culture medium and cultivated for 16 hours in an orbital shaker at 150 rpm and 25 °C in the dark.

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: .. 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates). .. After electroporation, cells were immediately resuspended in TSB to a final volume of 200 µl and recovered at 30 °C for 2 hr with shaking.

Electroporation:

Article Title: Construction of a Bile-responsive Expression System inLactobacillus plantarum
Article Snippet: .. This mixture was subjected to electroporation using GenePulser Xcell™ (BioRad, USA) using 200 Ω, 2.0 kV and 25 μF capacitance conditions. ..

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: .. Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: .. Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA). ..

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ]. .. Transfected parasites were selected and maintained in RPMI-1640 culture medium containing 100 μg/ml hygromycin and heterozygous promastigotes (LdASS +/+/Hyg ) cell lines were confirmed by PCR analysis.

Article Title: RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene
Article Snippet: .. Upon placing a concentrated cell suspension (300 μL containing 1 × 107 cells) in a previously chilled 0.4 cm-gap electroporation cuvette, plasmidic DNA (50 μg) was added, and the mixture was incubated on ice for 5 min. Next, cells were electroporated using the following conditions: 350 V, 1000 μF and 700 Ω in a GenePulser Xcell (Bio-rad, Hercules, CA, USA). ..

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: .. The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA). .. The parameters of electroporation were set as 2 kV, 50 μF, and 300 Ω. Electroporated cells were resuspended in 50 mL of culture medium and cultivated for 16 hours in an orbital shaker at 150 rpm and 25 °C in the dark.

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates). .. After electroporation, cells were immediately resuspended in TSB to a final volume of 200 µl and recovered at 30 °C for 2 hr with shaking.

Transfection:

Article Title: Phosphodiesterase beta is the master regulator of cAMP signalling during malaria parasite invasion
Article Snippet: Paragraph title: Transfection of P . falciparum blood stages ... The DNA-cytomix solution was added to 250 μL of the packed ring stage cultures at 5% to 10% parasitaemia and the sample electroporated at 950 μF capacitance and a voltage of 0.31 kV using a GenePulser Xcell (Bio-Rad).

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ). .. Transfected parasites were selected with 50 µg·ml−1 of hygromycin (Jena Bioscience) and then maintained with 100 µg·ml−1 .

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Paragraph title: Transfection of Cells ... Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms).

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: .. Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA). ..

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ]. .. Transfected parasites were selected and maintained in RPMI-1640 culture medium containing 100 μg/ml hygromycin and heterozygous promastigotes (LdASS +/+/Hyg ) cell lines were confirmed by PCR analysis.

Article Title: RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene
Article Snippet: Paragraph title: 2.4. Transfection and Selection of Giardia Trophozoites Containing the Vectors ... Upon placing a concentrated cell suspension (300 μL containing 1 × 107 cells) in a previously chilled 0.4 cm-gap electroporation cuvette, plasmidic DNA (50 μg) was added, and the mixture was incubated on ice for 5 min. Next, cells were electroporated using the following conditions: 350 V, 1000 μF and 700 Ω in a GenePulser Xcell (Bio-rad, Hercules, CA, USA).

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: .. All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA). .. Recombinants were expressed and purified by His GraviTrap (GE Healthcare) according to the manufacturer's protocol.

Article Title: On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei
Article Snippet: A total of 1 × 107 29.13.6 cells were used for each procyclic-form transfection. .. Then, 25 μg of linearized plasmid DNA was added to the solution and cells were pulsed twice at 1,600 V with a time constant of 0.6 ms on a GenePulser Xcell (BioRad, Hercules, CA, USA).

Cell Culture:

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA). .. The colonies resistant to zeocin were isolated and cultured for subsequent analysis.

Article Title: On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei
Article Snippet: Paragraph title: Cell culture ... Then, 25 μg of linearized plasmid DNA was added to the solution and cells were pulsed twice at 1,600 V with a time constant of 0.6 ms on a GenePulser Xcell (BioRad, Hercules, CA, USA).

Generated:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Polymerase Chain Reaction:

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: Every PCR-amplified sequence was sequenced at the VIB Genetic Service Facility (VIB Genetic Service Facility, Antwerp, Belgium). .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ).

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Both the 5’-flanking and 3’-flanking PCR products were cloned upstream and downstream of the hygromycin (HYG) or neomycin (NEO) drug cassette respectively, in pX-63-HYG and pX-63-NEO vectors. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Recombinant:

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: Paragraph title: Cloning, expression and purification of recombinant VTN-1 to -4 and -2-1 to -2-3 ... All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

DNA Extraction:

Article Title: Construction of a Bile-responsive Expression System inLactobacillus plantarum
Article Snippet: Paragraph title: DNA isolation, transformation, and bile induction ... This mixture was subjected to electroporation using GenePulser Xcell™ (BioRad, USA) using 200 Ω, 2.0 kV and 25 μF capacitance conditions.

Fluorescence:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Stable cell lines with intermediate levels of Sig-1R-GFP expression (determined by fluorescence microscopy) were identified and then maintained in DMEM supplemented with FCS (10%) and G418 (100 μg/mL).

Isolation:

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA). .. The colonies resistant to zeocin were isolated and cultured for subsequent analysis.

Subcloning:

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: All genes except the MRPA gene were PCR amplified from BPK282 DNA using Phusion high-fidelity polymerase (NEB) with overhang primers ( ) for subcloning in pGEMT (Promega) before final cloning in pLEXSY-hyg2. .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ).

Microscopy:

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Stable cell lines with intermediate levels of Sig-1R-GFP expression (determined by fluorescence microscopy) were identified and then maintained in DMEM supplemented with FCS (10%) and G418 (100 μg/mL).

Purification:

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Plasmids were linearized with the restriction endonuclease Hind III and Bgl II and the products (~2.72 kb for hygromycin cassette and 2.48 kb for neomycin cassette) containing both the flanking regions of LdASS and HYG or NEO drug selection marker were purified by agarose gel electrophoresis. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: Paragraph title: Cloning, expression and purification of recombinant VTN-1 to -4 and -2-1 to -2-3 ... All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

Sequencing:

Article Title: Molecular Preadaptation to Antimony Resistance in Leishmania donovani on the Indian Subcontinent
Article Snippet: Every PCR-amplified sequence was sequenced at the VIB Genetic Service Facility (VIB Genetic Service Facility, Antwerp, Belgium). .. Promastigotes of BPK026/0 cl5 were then transfected using GenePulser Xcell (Bio-Rad) as described previously ( ).

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: Atg7 knockdown by small interfering RNA (siRNA) transfection Atg7 and nonspecific sequence control (NS) siRNAs ON-TARGET plus smart pools were obtained from Dharmacon (Thermo Fisher Scientific Inc., Waltham, MA). .. Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA).

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Insertion of the 5’-flanking and 3’-flanking regions was confirmed by restriction digestion and DNA sequence analysis. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Small Interfering RNA:

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: Paragraph title: Atg7 knockdown by small interfering RNA (siRNA) transfection ... Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA).

CRISPR:

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates). .. For reverse CRISPR immunity assays targeting insertion vectors, additional plating in the presence of ATc at a final concentration of 0.5 µg/ml was performed in parallel using the same dilutions.

Plasmid Preparation:

Article Title: Phosphodiesterase beta is the master regulator of cAMP signalling during malaria parasite invasion
Article Snippet: A total of 80 μg of precipitated plasmid DNA was resuspended in 400 μL cytomix (120 mM KCl, 0.15 mM CaCl2 , 2 mM EGTA, 5 mM MgCl2 , 10 mM K2 HPO4 /KH2 PO4 , and 25 mM HEPES [pH 7]). .. The DNA-cytomix solution was added to 250 μL of the packed ring stage cultures at 5% to 10% parasitaemia and the sample electroporated at 950 μF capacitance and a voltage of 0.31 kV using a GenePulser Xcell (Bio-Rad).

Article Title: Construction of a Bile-responsive Expression System inLactobacillus plantarum
Article Snippet: On the other hand, 1 μg of plasmid DNA was added to 50 μL of ice-cold cell suspension (~109 CFU/mL) in a disposable cuvette (Gene Pulser® Cuvette, 0.2 cm electrode gap; Bio-Rad, Hercules, CA, USA) and held on ice for 5 min. .. This mixture was subjected to electroporation using GenePulser Xcell™ (BioRad, USA) using 200 Ω, 2.0 kV and 25 μF capacitance conditions.

Article Title: A non-catalytic role of RecBCD in homology directed gap repair and translesion synthesis
Article Snippet: .. Monitoring HDGR and TLS To 40 μl aliquot of competent cells, prepared as previously described , 1 ng of the lesion-carrying vector mixed with 1 ng of the internal standard (pVP146) was added and electroporated in a GenePulser Xcell from BioRad (2.5 kV, 25 μF, 200 Ω). ..

Article Title: Choline Is an Intracellular Messenger Linking Extracellular Stimuli to IP3-Evoked Ca2+ Signals through Sigma-1 Receptors
Article Snippet: .. Cells (500 μL) were transferred to electroporation cuvettes (800 μL, 4-mm gap; Eppendorf, Hamburg, Germany) with plasmid DNA (5-10 μg/cuvette) and the cells were subjected to electroporation using a GenePulser Xcell (BioRad, 200-250V, 700-900 μF, 18-20 ms). .. Transfected cells were plated in Opti-MEM I in 6-well plates, FBS (10%) was added after 4 h, and the medium was replaced after 24 h. Neuro-2A cells stably expressing Sig-1R-GFP were generated by transfecting cells with plasmid encoding human Sig-1R-GFP using Lipofectamine.

Article Title: Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization
Article Snippet: .. After construction, 86676_3HA_Pur or 16263_3HA_Pur were linearized for homologous recombination by PstI or NsiI, respectively and ~10 µg of plasmid was electroporated into 300 µl of Giardia in a 0.4 cm cuvette using a GenePulser XCell (BioRad) set to 375 V, 1000 µF and 750 ohms. ..

Article Title: Consequences of Cas9 cleavage in the chromosome of Escherichia coli
Article Snippet: Mu Gam Electrocompetent cells of the E. coli strain MG1655 carrying plasmid pLC13 were prepared. .. The pCas9, pCas9::LacZ1 and pCas9::LacZ2 plasmids (40 ng each) were transformed into electrocompetent cells using 1 mm gap cuvette and the GenePulser Xcell (Bio-Rad) set to 1800 V, 25 uF, 200 Ω.

Article Title: Illustrating and Enhancing the Biosynthesis of Astaxanthin and Docosahexaenoic Acid in Aurantiochytrium sp. SK4
Article Snippet: Paragraph title: 3.8. Construction of Plasmid p-VBIG and Transformation ... The schematic diagram of p-VBIG is shown in A. Aurantiochytrium sp. SK4 was transformed with p-VBIG by electroporation using a GenePulser Xcell™ (Bio-Rad, Hercules, CA, USA).

Article Title: Molecular Camouflage of Plasmodium falciparum Merozoites by Binding of Host Vitronectin to P47 Fragment of SERA5
Article Snippet: The amplified fragments were digested and ligated in-frame into the Eco RI and Not I sites of the pPICZα A expression vector for Pichia pastoris Expression System (Thermo Fisher Scientific). .. All constructs were linearized by Pme I digestion and then transfected to GS115 strain using GenePulser Xcell (Bio-Rad, Hercules, CA, USA).

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: Transformation assays S. aureus RN4220 plasmid preps were dialyzed on 0.025 µm nitrocellulose filters (Millipore) and then quantified using a NanoDrop 2000c Spectrophotometer (Thermo Scientific). .. 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates).

Article Title: On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei
Article Snippet: .. Then, 25 μg of linearized plasmid DNA was added to the solution and cells were pulsed twice at 1,600 V with a time constant of 0.6 ms on a GenePulser Xcell (BioRad, Hercules, CA, USA). ..

Selection:

Article Title: Phosphodiesterase beta is the master regulator of cAMP signalling during malaria parasite invasion
Article Snippet: The DNA-cytomix solution was added to 250 μL of the packed ring stage cultures at 5% to 10% parasitaemia and the sample electroporated at 950 μF capacitance and a voltage of 0.31 kV using a GenePulser Xcell (Bio-Rad). .. Twenty-four hours later, the selection drug WR99210 was added at 5 nM concentration.

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Plasmids were linearized with the restriction endonuclease Hind III and Bgl II and the products (~2.72 kb for hygromycin cassette and 2.48 kb for neomycin cassette) containing both the flanking regions of LdASS and HYG or NEO drug selection marker were purified by agarose gel electrophoresis. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Article Title: RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene
Article Snippet: Paragraph title: 2.4. Transfection and Selection of Giardia Trophozoites Containing the Vectors ... Upon placing a concentrated cell suspension (300 μL containing 1 × 107 cells) in a previously chilled 0.4 cm-gap electroporation cuvette, plasmidic DNA (50 μg) was added, and the mixture was incubated on ice for 5 min. Next, cells were electroporated using the following conditions: 350 V, 1000 μF and 700 Ω in a GenePulser Xcell (Bio-rad, Hercules, CA, USA).

Agarose Gel Electrophoresis:

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Plasmids were linearized with the restriction endonuclease Hind III and Bgl II and the products (~2.72 kb for hygromycin cassette and 2.48 kb for neomycin cassette) containing both the flanking regions of LdASS and HYG or NEO drug selection marker were purified by agarose gel electrophoresis. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Spectrophotometry:

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: Transformation assays S. aureus RN4220 plasmid preps were dialyzed on 0.025 µm nitrocellulose filters (Millipore) and then quantified using a NanoDrop 2000c Spectrophotometer (Thermo Scientific). .. 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates).

Concentration Assay:

Article Title: Phosphodiesterase beta is the master regulator of cAMP signalling during malaria parasite invasion
Article Snippet: The DNA-cytomix solution was added to 250 μL of the packed ring stage cultures at 5% to 10% parasitaemia and the sample electroporated at 950 μF capacitance and a voltage of 0.31 kV using a GenePulser Xcell (Bio-Rad). .. Twenty-four hours later, the selection drug WR99210 was added at 5 nM concentration.

Article Title: Elevated autophagy gene expression in adipose tissue of obese humans: A potential non-cell-cycle-dependent function of E2F1
Article Snippet: .. Each siRNA was transfected into differentiated epididymal preadipocyte cell line at a concentration of 1 nmol/well by electroporation with GenePulser Xcell (Bio-Rad Laboratories Inc., Hercules, CA). ..

Article Title: Conditional tolerance of temperate phages via transcription-dependent CRISPR-Cas targeting
Article Snippet: 50 µl aliquots of electrocompetent cells were transformed in triplicate with 80 ng dialyzed DNA per transformation using a GenePulser Xcell (BioRad) with the following parameters: 2900 V, 25 µF, 100 Ω, 2 mm (technical replicates). .. For reverse CRISPR immunity assays targeting insertion vectors, additional plating in the presence of ATc at a final concentration of 0.5 µg/ml was performed in parallel using the same dilutions.

Article Title: On the extent and role of the small proteome in the parasitic eukaryote Trypanosoma brucei
Article Snippet: Then, 25 μg of linearized plasmid DNA was added to the solution and cells were pulsed twice at 1,600 V with a time constant of 0.6 ms on a GenePulser Xcell (BioRad, Hercules, CA, USA). .. Phleomycin and blasticidin were added to a final concentration of 2.5 μg/ml and 10 μg/ml, respectively.

Marker:

Article Title: Genetic Manipulation of Leishmania donovani to Explore the Involvement of Argininosuccinate Synthase in Oxidative Stress Management
Article Snippet: Plasmids were linearized with the restriction endonuclease Hind III and Bgl II and the products (~2.72 kb for hygromycin cassette and 2.48 kb for neomycin cassette) containing both the flanking regions of LdASS and HYG or NEO drug selection marker were purified by agarose gel electrophoresis. .. Wild-type (WT, LdASS +/+/+ ) L . donovani promastigotes were transfected via electroporation with 5 μg of the linearized fragment (5’-LdASS -HYG-3’-LdASS ) using a GenePulser XCell (Bio-Rad) system as previously described [ ].

Homologous Recombination:

Article Title: Hybrid Structured Illumination Expansion Microscopy Reveals Microbial Cytoskeleton Organization
Article Snippet: .. After construction, 86676_3HA_Pur or 16263_3HA_Pur were linearized for homologous recombination by PstI or NsiI, respectively and ~10 µg of plasmid was electroporated into 300 µl of Giardia in a 0.4 cm cuvette using a GenePulser XCell (BioRad) set to 375 V, 1000 µF and 750 ohms. ..

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