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Thermo Fisher bsrgi
Bsrgi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 7 article reviews
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bsrgi - by Bioz Stars, 2020-01
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Clone Assay:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: AAV-CAG-ChR2-GFP (Addgene plasmid #26929) was a gift from Edward Boyden, and was used as template for cloning GFP. .. AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher).

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: Then, the 2A-mCherry sequence (see above) was introduced into BshVI/SalI-digested pAAV-hSyn-Cacna2d4 using the In-Fusion HD Cloning kit (Takara Bio) with forward 5′-aactcctgcggatcggagtgaaacagactttgaattttgaccttct-3′ and reverse 5′-gcttctgcaggtcgatcacttgtacagctcg-3′ primers. pLenti-hSyn-Cacna2d1-HA was made using a pLenti-vector of the second generation (pLenti-synapsin-ChR2; provided by Thomas Oertner and Tobias Rose; FMI Basel). .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: The 5’ and 3’UTRs were cloned by RACE (Rapid Amplification of cDNA ends) with primers in TABLE SI using 5’ and 3’ RACE Systems (Invitrogen) according to manufacturer’s instructions for first strand cDNA synthesis, and the Expand High Fidelity PCR System (Roche) for PCRs. .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: Paragraph title: Plasmids and cloning. ... The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing.

Amplification:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher). .. By sequential PCR amplification, mCherry alone, Zip14-IRES-GFP amplicons, containing N-terminal BamHI and C-terminal BsrGI digestion sites, were introduced into digested AAV-CAG backbone to get the AAV vectors.

Titration:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: Paragraph title: Virus production, purification and titration ... AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher).

Synthesized:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: To generate cDNAs spanning exons 1–35 including all but 199 bases of the 5’ end and 46 bases of the 3’ end of the ORF by long-range RT-PCR, AccuPrime™ High Fidelity Taq DNA Polymerase (Invitrogen) and primers in TABLE SI were used to amplify either oligo-dT primed first strand cDNA from adults, or a mixture of oligo-dT primed and random hexamer primed first strand cDNA from 24 hpf embryos synthesized with a SuperScript™ II First Strand Synthesis kit (Invitrogen). .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Construct:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: Virus production, purification and titration For adeno-associated virus production, we constructed two different AAV vectors, including AAV-CAG-Zip14-IRES-GFP and AAV-CAG-mCherry. pAAV-Ef1a-mCherry- IRES-Cre (Addgene plasmid #55632) was a gift from Karl Deisseroth, and was used as PCR template for cloning mCherry and IRES. .. AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher).

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: Next, the zinc-binding domain of Egr1 was deleted by digestion and subsequent ligation of the pKan-CMV-Egr1-2A-mCherry construct with XmnI. .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: NR4A2 Is Regulated by Gastrin and Influences Cellular Responses of Gastric Adenocarcinoma Cells
Article Snippet: .. Zfp36l1 (Berg36) entry clone (Berg36 ORF Express Shuttle Clone) was ordered from GeneCopoeia (USA), and pZfp36l1-DEST was constructed by homologous recombination with entry clone and pDEST26 (Invitrogen). pCONTROL vectors were constructed by restriction cutting in att sites in pEF5/FRT/V5-DEST with EcoRV, and in pDEST26 with BsrGI, followed by re-ligation of att sites, thereby removing the insert between the att sites. pEF5/FRT/V5/GW-CAT was purchased from Invitrogen. .. Transient transfection and gastrin treatment of cells AGS-GR cells (5.0 x 105 /well) were plated in 6-well plates and transfected after 24 h with 2.5 µg plasmid and 12.5 µl Metafectene PRO transfection reagent (Biontex Laboratories GmbH, Martinsried, Germany) per well.

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Incubation:

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing. .. A plasmid expressing M1 fused to the red fluorescent protein mCardinal (Card-M1) was obtained by cloning the M1 insert (influenza A/FPV/Rostock/34 virus [ ]) into the mCardinal-C1 backbone vector.

Expressing:

Article Title: NR4A2 Is Regulated by Gastrin and Influences Cellular Responses of Gastric Adenocarcinoma Cells
Article Snippet: Paragraph title: Expression plasmids ... Zfp36l1 (Berg36) entry clone (Berg36 ORF Express Shuttle Clone) was ordered from GeneCopoeia (USA), and pZfp36l1-DEST was constructed by homologous recombination with entry clone and pDEST26 (Invitrogen). pCONTROL vectors were constructed by restriction cutting in att sites in pEF5/FRT/V5-DEST with EcoRV, and in pDEST26 with BsrGI, followed by re-ligation of att sites, thereby removing the insert between the att sites. pEF5/FRT/V5/GW-CAT was purchased from Invitrogen.

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: A plasmid expressing a myristoylated-palmitoylated yellow fluorescent protein (YFP) ( , ) was modified to obtain a myristoylated-palmitoylated mRFP (myr-palm mRFP) plasmid. .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing.

Modification:

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: A plasmid expressing a myristoylated-palmitoylated yellow fluorescent protein (YFP) ( , ) was modified to obtain a myristoylated-palmitoylated mRFP (myr-palm mRFP) plasmid. .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing.

Derivative Assay:

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: .. This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Transfection:

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. This construct was then mixed with packaging plasmids (psPAX2 and pMD2.G, Addgene plasmids 12260 and 12259, respectively) and used with Lentiphos HT transfection system (ClonTech, Palo Alto, CA) to transfect 293T cells (50–60% confluency) for production of VSV-G pseudotyped, replication-incompetent lenti-virus.

Ligation:

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: Next, the zinc-binding domain of Egr1 was deleted by digestion and subsequent ligation of the pKan-CMV-Egr1-2A-mCherry construct with XmnI. .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen). .. A cDNA spanning the 5’UTR through the 3’UTR was generated from random hexamer primed cDNAs by long-distance RT-PCR using AccuPrime™ High Fidelity Taq DNA Polymerase (Invitrogen), subcloned into pCR-XL-TOPO (Invitrogen) and sequenced.

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing. .. A plasmid expressing M1 fused to the red fluorescent protein mCardinal (Card-M1) was obtained by cloning the M1 insert (influenza A/FPV/Rostock/34 virus [ ]) into the mCardinal-C1 backbone vector.

Generated:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: Products were subcloned into pCR-XL-TOPO (Invitrogen) and a sequence contig generated. .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Polymerase Chain Reaction:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: Virus production, purification and titration For adeno-associated virus production, we constructed two different AAV vectors, including AAV-CAG-Zip14-IRES-GFP and AAV-CAG-mCherry. pAAV-Ef1a-mCherry- IRES-Cre (Addgene plasmid #55632) was a gift from Karl Deisseroth, and was used as PCR template for cloning mCherry and IRES. .. AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher).

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen). .. A cDNA spanning the 5’UTR through the 3’UTR was generated from random hexamer primed cDNAs by long-distance RT-PCR using AccuPrime™ High Fidelity Taq DNA Polymerase (Invitrogen), subcloned into pCR-XL-TOPO (Invitrogen) and sequenced.

DNA Sequencing:

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Sequencing:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher). .. The AAV constructs were confirmed by sequencing, and then co-transfected with pDeltaF6 and AAV 2/9 Helper plasmids, in a ratio of 1:2:1.6, into 293T cells by calcium phosphate.

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: Then, the 2A-mCherry sequence (see above) was introduced into BshVI/SalI-digested pAAV-hSyn-Cacna2d4 using the In-Fusion HD Cloning kit (Takara Bio) with forward 5′-aactcctgcggatcggagtgaaacagactttgaattttgaccttct-3′ and reverse 5′-gcttctgcaggtcgatcacttgtacagctcg-3′ primers. pLenti-hSyn-Cacna2d1-HA was made using a pLenti-vector of the second generation (pLenti-synapsin-ChR2; provided by Thomas Oertner and Tobias Rose; FMI Basel). .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: Products were subcloned into pCR-XL-TOPO (Invitrogen) and a sequence contig generated. .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Article Title: NR4A2 Is Regulated by Gastrin and Influences Cellular Responses of Gastric Adenocarcinoma Cells
Article Snippet: Sequence verification of the plasmid identified -128 to +154 of the NR4A2 promoter, including the CRE element. pCONTROL-luc were obtained from Panomics (CA, USA). .. Zfp36l1 (Berg36) entry clone (Berg36 ORF Express Shuttle Clone) was ordered from GeneCopoeia (USA), and pZfp36l1-DEST was constructed by homologous recombination with entry clone and pDEST26 (Invitrogen). pCONTROL vectors were constructed by restriction cutting in att sites in pEF5/FRT/V5-DEST with EcoRV, and in pDEST26 with BsrGI, followed by re-ligation of att sites, thereby removing the insert between the att sites. pEF5/FRT/V5/GW-CAT was purchased from Invitrogen.

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing. .. A plasmid expressing M1 fused to the red fluorescent protein mCardinal (Card-M1) was obtained by cloning the M1 insert (influenza A/FPV/Rostock/34 virus [ ]) into the mCardinal-C1 backbone vector.

Nucleic Acid Electrophoresis:

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Subcloning:

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: .. This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Purification:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: Paragraph title: Virus production, purification and titration ... AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher).

Reverse Transcription Polymerase Chain Reaction:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: To generate cDNAs spanning exons 1–35 including all but 199 bases of the 5’ end and 46 bases of the 3’ end of the ORF by long-range RT-PCR, AccuPrime™ High Fidelity Taq DNA Polymerase (Invitrogen) and primers in TABLE SI were used to amplify either oligo-dT primed first strand cDNA from adults, or a mixture of oligo-dT primed and random hexamer primed first strand cDNA from 24 hpf embryos synthesized with a SuperScript™ II First Strand Synthesis kit (Invitrogen). .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Rapid Amplification of cDNA Ends:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: The 5’ and 3’UTRs were cloned by RACE (Rapid Amplification of cDNA ends) with primers in TABLE SI using 5’ and 3’ RACE Systems (Invitrogen) according to manufacturer’s instructions for first strand cDNA synthesis, and the Expand High Fidelity PCR System (Roche) for PCRs. .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Plasmid Preparation:

Article Title: Metastatic cancers promote cachexia through altered zinc homeostasis in skeletal muscle
Article Snippet: .. AAV-CAG-ChR2-GFP was also used as backbone AAV vector with CAG promoter after digesting by BamHI (Roche) and BsrGI (Thermo Fisher). .. By sequential PCR amplification, mCherry alone, Zip14-IRES-GFP amplicons, containing N-terminal BamHI and C-terminal BsrGI digestion sites, were introduced into digested AAV-CAG backbone to get the AAV vectors.

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: The pAAV-hSyn-Cacna2d4-2A-mCherry plasmid was produced by first introducing the human Cacna2d4 gene (kind gift from Dr. F. Haeseleer, University of Washington) ( ) into pAAV-hSyn-MCS using AsiSI/ClaI. .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: NR4A2 Is Regulated by Gastrin and Influences Cellular Responses of Gastric Adenocarcinoma Cells
Article Snippet: NR4A2-EGFP was a kind gift from Prof. Evely Murphy, University College Dublin, Ireland [ ]. pICER IIy and pICER I were constructed via homologous recombination of ICER IIy or ICER I containing pDONR201 plasmid [ ] and pEF5/FRT/V5-DEST (Invitrogen). .. Zfp36l1 (Berg36) entry clone (Berg36 ORF Express Shuttle Clone) was ordered from GeneCopoeia (USA), and pZfp36l1-DEST was constructed by homologous recombination with entry clone and pDEST26 (Invitrogen). pCONTROL vectors were constructed by restriction cutting in att sites in pEF5/FRT/V5-DEST with EcoRV, and in pDEST26 with BsrGI, followed by re-ligation of att sites, thereby removing the insert between the att sites. pEF5/FRT/V5/GW-CAT was purchased from Invitrogen.

Article Title: Phosphatidylserine Lateral Organization Influences the Interaction of Influenza Virus Matrix Protein 1 with Lipid Membranes
Article Snippet: A plasmid expressing a myristoylated-palmitoylated yellow fluorescent protein (YFP) ( , ) was modified to obtain a myristoylated-palmitoylated mRFP (myr-palm mRFP) plasmid. .. The mRFP-Lact-C2 and myristoylated-palmitoylated YFP plasmids were incubated with the restriction enzymes AgeI and BsrgI (Thermo Fisher Scientific, Darmstadt, Germany); the two products were then ligated overnight with the T4 DNA ligase enzyme (Thermo Fisher Scientific, Darmstadt, Germany), and ligation success was confirmed by sequencing.

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: .. This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Dominant Negative Mutation:

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: Using this strategy, a 282 bp fragment is removed, resulting in a dominant-negative version of the Egr1 gene (Egr1dN; ( ). .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Random Hexamer Labeling:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: To generate cDNAs spanning exons 1–35 including all but 199 bases of the 5’ end and 46 bases of the 3’ end of the ORF by long-range RT-PCR, AccuPrime™ High Fidelity Taq DNA Polymerase (Invitrogen) and primers in TABLE SI were used to amplify either oligo-dT primed first strand cDNA from adults, or a mixture of oligo-dT primed and random hexamer primed first strand cDNA from 24 hpf embryos synthesized with a SuperScript™ II First Strand Synthesis kit (Invitrogen). .. A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Produced:

Article Title: Calcium Channel Subunit α2δ4 Is Regulated by Early Growth Response 1 and Facilitates Epileptogenesis
Article Snippet: The pAAV-hSyn-Cacna2d4-2A-mCherry plasmid was produced by first introducing the human Cacna2d4 gene (kind gift from Dr. F. Haeseleer, University of Washington) ( ) into pAAV-hSyn-MCS using AsiSI/ClaI. .. The ChR2 insert was removed by digestion with AgeI and BsrGI, and sticky ends were afterward filled up using Klenow Fragment (Thermo Fisher Scientific).

Article Title: Morphology of Axonal Projections From the High Vocal Center to Vocal Motor Cortex in Songbirds
Article Snippet: .. This vector was produced by removing the CAG-GFP cassette from pCAG-GFP ( ; Addgene plasmid 11150) using SpeI and BsrGI, and subcloning this cassette into the lentiviral backbone, derived from FCK-ChR2-GFP (Addgene plasmid 15814; ) cut with XbaI and BsrGI (all enzymes from Fermentas, Burlington, ON, Canada). .. The lentiviral vector construct was verified by enzymatic digestion, gel electrophoresis, and DNA sequencing (DNA Core Facility, USC).

Molecular Cloning:

Article Title: mll ortholog containing functional domains of human MLL is expressed throughout the zebrafish lifespan and in haematopoietic tissues
Article Snippet: Paragraph title: Molecular cloning of zebrafish mll cDNAs ... A full-length ORF cDNA subclone from 24 hpf embryos was obtained by restriction enzyme cleavage of the 5’UTR-exon 3 PCR product with NotI and BsrGI, of the larger central exon 1–35 subclone with BsrGI and ClaI, and of the exon 33–35 PCR product with ClaI and KpnI, followed by ligation of the 5’ and 3’ fragments to the BsrGI or ClaI sites in the larger central fragment and to NotI and KpnI sites in pCR-XL-TOPO (Invitrogen).

Homologous Recombination:

Article Title: NR4A2 Is Regulated by Gastrin and Influences Cellular Responses of Gastric Adenocarcinoma Cells
Article Snippet: .. Zfp36l1 (Berg36) entry clone (Berg36 ORF Express Shuttle Clone) was ordered from GeneCopoeia (USA), and pZfp36l1-DEST was constructed by homologous recombination with entry clone and pDEST26 (Invitrogen). pCONTROL vectors were constructed by restriction cutting in att sites in pEF5/FRT/V5-DEST with EcoRV, and in pDEST26 with BsrGI, followed by re-ligation of att sites, thereby removing the insert between the att sites. pEF5/FRT/V5/GW-CAT was purchased from Invitrogen. .. Transient transfection and gastrin treatment of cells AGS-GR cells (5.0 x 105 /well) were plated in 6-well plates and transfected after 24 h with 2.5 µg plasmid and 12.5 µl Metafectene PRO transfection reagent (Biontex Laboratories GmbH, Martinsried, Germany) per well.

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    Thermo Fisher bsp1407i
    Bsp1407i, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsp1407i/product/Thermo Fisher
    Average 87 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsp1407i - by Bioz Stars, 2020-01
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    99
    Thermo Fisher bsrgi
    Bsrgi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsrgi/product/Thermo Fisher
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    bsrgi - by Bioz Stars, 2020-01
    99/100 stars
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