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TaKaRa la taq polymerase
La Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 187 article reviews
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la taq polymerase - by Bioz Stars, 2020-04
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Diagnostic Assay:

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; ) and CVB5 (Faulkner; ) (Fig. ). .. Takara LA was the only polymerase other than KlenTaq LA to successfully amplify near full-length genome products of ~8 kb, and this enzyme was used in further testing of the method using four stool specimens from the NSW Health Pathology East virology diagnostic laboratory, which were previously confirmed as EV-positive using diagnostic qPCR.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; JX312064.1) and CVB5 (Faulkner; AF114383.1) (Fig. ). .. Takara LA was the only polymerase other than KlenTaq LA to successfully amplify near full-length genome products of ~8 kb, and this enzyme was used in further testing of the method using four stool specimens from the NSW Health Pathology East virology diagnostic laboratory, which were previously confirmed as EV-positive using diagnostic qPCR.

Clone Assay:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. After verification by sequencing (Genomics), the fragment was cloned into the pCAMBIA1300 binary vector (Cambia) and subsequently introduced into homozygous cdkg1 plants.

Amplification:

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: .. Genotypic characterisation of exopolysaccharides The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: .. The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; ) and CVB5 (Faulkner; ) (Fig. ). .. As anticipated, near-full length EV amplicons were successfully amplified from all four specimens (Fig. ).

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. Animals were injected with the purified PCR products at 1–10 ng/µl together with the co-injection marker pRF4 (rol-6(su1006) ) at 100 ng/µl and analyzed for rescue of IDA-1::GFP intensity and distribution.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: Paragraph title: Comparison of Polymerases for Efficient Near Full-Length EV Amplification ... The following enzymes were tested: Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech).

Article Title: Noninvasive Immunohistochemical Diagnosis and Novel MUC1 Mutations Causing Autosomal Dominant Tubulointerstitial Kidney Disease
Article Snippet: .. These sequences were then read with the Illumina system (see the following references for an explanation of the Illumina system)., The VNTR region was directly amplified from genomic DNA using primers PS2F-T7 (5′-GGATCCTAATACGACTCACTATAGGAACAGACCACCATGGGAGAAAAGGAGACTTCGGCTACCCAG-3′) and PS3 (specified above) and long-range PCR (TaKaRa LA Taq DNA Polymerase with GC Buffers; Takara, Mountain View, CA). .. The resulting PCR products were purified using AmpureXP Beads fragmented to 400 bp with the Covaris E220 System and purified again with AmpureXP Beads.

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: .. For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. After verification by sequencing (Genomics), the fragment was cloned into the pCAMBIA1300 binary vector (Cambia) and subsequently introduced into homozygous cdkg1 plants.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; JX312064.1) and CVB5 (Faulkner; AF114383.1) (Fig. ). .. As anticipated, near-full length EV amplicons were successfully amplified from all four specimens (Fig. ).

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: .. The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Polymerase Chain Reaction:

Article Title: Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products
Article Snippet: Paragraph title: PCR ... The same results were obtained by the other polymerase systems: LA Taq polymerase (Takara-bio), Tth polymerase (Applied-boisystems); Expand High Fidelity, Expand High FidelityPLUS and Expand Long Template polymerase (Roche-diagnostics); TITANIUM Taq polymerase (Becton-Dickinson-Clontech); and Taq polymerase (Promega).

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: .. Genotypic characterisation of exopolysaccharides The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: Paragraph title: Optimisation of Near Full-Length Genome PCR ... In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; ) and CVB5 (Faulkner; ) (Fig. ).

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: To confirm the gene location of gv560 , small PCR fragments (1 kb in size) covering the ∼2-kb promoter region, entire coding region, and intron-exon junction regions of pag-3 were generated for sequencing (ABI 373 DNA sequencer). .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI).

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: This was used as a template for full-length EV amplification PCR, to test the efficiency of three commercially available long amplifying polymerases in comparison to KlenTaq LA (Clontech), which discontinued production in late 2015 . .. The following enzymes were tested: Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech).

Article Title: Molecular Characterization Reveals Three Distinct Clonal Groups among Clinical Shiga Toxin-Producing Escherichia coli Strains of Serogroup O103
Article Snippet: .. Long-range PCR screenings were performed by using TaKaRa LA Taq polymerase (TaKaRa Bio, Inc., Ohtsu, Japan). .. The O103-antigen biosynthesis gene cluster and its flanking regions were amplified using a PCR primer pair, O55re-1F and O55re-1R ( ).

Article Title: Noninvasive Immunohistochemical Diagnosis and Novel MUC1 Mutations Causing Autosomal Dominant Tubulointerstitial Kidney Disease
Article Snippet: .. These sequences were then read with the Illumina system (see the following references for an explanation of the Illumina system)., The VNTR region was directly amplified from genomic DNA using primers PS2F-T7 (5′-GGATCCTAATACGACTCACTATAGGAACAGACCACCATGGGAGAAAAGGAGACTTCGGCTACCCAG-3′) and PS3 (specified above) and long-range PCR (TaKaRa LA Taq DNA Polymerase with GC Buffers; Takara, Mountain View, CA). .. The resulting PCR products were purified using AmpureXP Beads fragmented to 400 bp with the Covaris E220 System and purified again with AmpureXP Beads.

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. As CHC-R primer was designed 180 bp downstream of CMP-R primer, PCR with the CHC-F/CHC-R primer set was not able to amplify a 1.7-kb fragment in homozygous plants even if the complementation fragment was integrated into the genome.

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: .. The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Real-time Polymerase Chain Reaction:

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; ) and CVB5 (Faulkner; ) (Fig. ). .. Takara LA was the only polymerase other than KlenTaq LA to successfully amplify near full-length genome products of ~8 kb, and this enzyme was used in further testing of the method using four stool specimens from the NSW Health Pathology East virology diagnostic laboratory, which were previously confirmed as EV-positive using diagnostic qPCR.

Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples
Article Snippet: In search of an alternative enzyme, three different long amplifying polymerases, including the Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech) were compared against KlenTaq for their efficiency in amplifying two American Type Culture Collection (ATCC) EV prototype strains, CVB3 (Nancy; JX312064.1) and CVB5 (Faulkner; AF114383.1) (Fig. ). .. Takara LA was the only polymerase other than KlenTaq LA to successfully amplify near full-length genome products of ~8 kb, and this enzyme was used in further testing of the method using four stool specimens from the NSW Health Pathology East virology diagnostic laboratory, which were previously confirmed as EV-positive using diagnostic qPCR.

Expressing:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. To determine whether the effect of pag-3 mutants on IDA-1::GFP expression is a cell autonomous, the full-length cDNA of pag-3 was specifically expressed in a limited number of neurons (including VC4 and VC5) driven by the 3-kb promoter of cat-1 to generate Pcat-1 PAG-3 in the PCRII-TOPO vector that was confirmed by sequencing.

Derivative Assay:

Article Title: Noninvasive Immunohistochemical Diagnosis and Novel MUC1 Mutations Causing Autosomal Dominant Tubulointerstitial Kidney Disease
Article Snippet: These sequences were then read with the Illumina system (see the following references for an explanation of the Illumina system)., The VNTR region was directly amplified from genomic DNA using primers PS2F-T7 (5′-GGATCCTAATACGACTCACTATAGGAACAGACCACCATGGGAGAAAAGGAGACTTCGGCTACCCAG-3′) and PS3 (specified above) and long-range PCR (TaKaRa LA Taq DNA Polymerase with GC Buffers; Takara, Mountain View, CA). .. In this process, the individual nucleotide sequences derived from the PCR are sequenced in parallel, resulting in multiple, repeated sequence readings from the VNTR.

Electron Microscopy:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For scanning electron microscopy examination, the opened flowers of the wild type and cdkg1 were dissected and the dehiscent anthers were coated with 8-nm gold. .. For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: Paragraph title: RT-PCR ... The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan).

Generated:

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The first-strand cDNA was generated from 4 µg of total RNA of each using Ready-To-Go You-Prime First-Strand Beads (GE Healthcare, Buckinghamshire, UK) with an oligo dT primer. .. The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan).

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: To confirm the gene location of gv560 , small PCR fragments (1 kb in size) covering the ∼2-kb promoter region, entire coding region, and intron-exon junction regions of pag-3 were generated for sequencing (ABI 373 DNA sequencer). .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI).

Transmission Assay:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: Sections were observed on an H-600 transmission electron microscope (Hitachi). .. For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R.

Sequencing:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: To confirm the gene location of gv560 , small PCR fragments (1 kb in size) covering the ∼2-kb promoter region, entire coding region, and intron-exon junction regions of pag-3 were generated for sequencing (ABI 373 DNA sequencer). .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI).

Article Title: Noninvasive Immunohistochemical Diagnosis and Novel MUC1 Mutations Causing Autosomal Dominant Tubulointerstitial Kidney Disease
Article Snippet: Paragraph title: Illumina Sequencing of the VNTR Region of MUC1 ... These sequences were then read with the Illumina system (see the following references for an explanation of the Illumina system)., The VNTR region was directly amplified from genomic DNA using primers PS2F-T7 (5′-GGATCCTAATACGACTCACTATAGGAACAGACCACCATGGGAGAAAAGGAGACTTCGGCTACCCAG-3′) and PS3 (specified above) and long-range PCR (TaKaRa LA Taq DNA Polymerase with GC Buffers; Takara, Mountain View, CA).

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. After verification by sequencing (Genomics), the fragment was cloned into the pCAMBIA1300 binary vector (Cambia) and subsequently introduced into homozygous cdkg1 plants.

Injection:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. Animals were injected with the purified PCR products at 1–10 ng/µl together with the co-injection marker pRF4 (rol-6(su1006) ) at 100 ng/µl and analyzed for rescue of IDA-1::GFP intensity and distribution.

Fluorescence:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: The quenching time was recorded when the callose fluorescence of each tetrad disappeared. .. For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R.

Mutagenesis:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. Animals were injected with the purified PCR products at 1–10 ng/µl together with the co-injection marker pRF4 (rol-6(su1006) ) at 100 ng/µl and analyzed for rescue of IDA-1::GFP intensity and distribution.

Microscopy:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: Paragraph title: Phenotype Characterization and Microscopy ... For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R.

Purification:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. Animals were injected with the purified PCR products at 1–10 ng/µl together with the co-injection marker pRF4 (rol-6(su1006) ) at 100 ng/µl and analyzed for rescue of IDA-1::GFP intensity and distribution.

Article Title: Noninvasive Immunohistochemical Diagnosis and Novel MUC1 Mutations Causing Autosomal Dominant Tubulointerstitial Kidney Disease
Article Snippet: These sequences were then read with the Illumina system (see the following references for an explanation of the Illumina system)., The VNTR region was directly amplified from genomic DNA using primers PS2F-T7 (5′-GGATCCTAATACGACTCACTATAGGAACAGACCACCATGGGAGAAAAGGAGACTTCGGCTACCCAG-3′) and PS3 (specified above) and long-range PCR (TaKaRa LA Taq DNA Polymerase with GC Buffers; Takara, Mountain View, CA). .. The resulting PCR products were purified using AmpureXP Beads fragmented to 400 bp with the Covaris E220 System and purified again with AmpureXP Beads.

Transgenic Assay:

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. For cdkg1 background verification, SC-F/SC-R primers were used to validate DNA deletion of CDKG1 for the transformants, LP/RP primers were used to detect either CDKG1 genomic sequence or transgenic complementation fragment, and the genomic-specific primers CHC-F/CHC-R were used to validate the homozygous cdkg1 background.

Chloramphenicol Acetyltransferase Assay:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. To determine whether the effect of pag-3 mutants on IDA-1::GFP expression is a cell autonomous, the full-length cDNA of pag-3 was specifically expressed in a limited number of neurons (including VC4 and VC5) driven by the 3-kb promoter of cat-1 to generate Pcat-1 PAG-3 in the PCRII-TOPO vector that was confirmed by sequencing.

Plasmid Preparation:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI). .. To determine whether the effect of pag-3 mutants on IDA-1::GFP expression is a cell autonomous, the full-length cDNA of pag-3 was specifically expressed in a limited number of neurons (including VC4 and VC5) driven by the 3-kb promoter of cat-1 to generate Pcat-1 PAG-3 in the PCRII-TOPO vector that was confirmed by sequencing.

Article Title: CYCLIN-DEPENDENT KINASE G1 Is Associated with the Spliceosome to Regulate CALLOSE SYNTHASE5 Splicing and Pollen Wall Formation in Arabidopsis
Article Snippet: For cdkg1 complementation, the 3.6-kb CDKG1 genomic fragment was amplified using LA-Taq polymerase (Takara) with primer set CMP-F/CMP-R. .. After verification by sequencing (Genomics), the fragment was cloned into the pCAMBIA1300 binary vector (Cambia) and subsequently introduced into homozygous cdkg1 plants.

Agarose Gel Electrophoresis:

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: Genotypic characterisation of exopolysaccharides The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: Detection of virulence factors of South African Lactococcus garvieae isolated from rainbow trout, Oncorhynchus mykiss (Walbaum)
Article Snippet: The L. garvieae EPS synthesis gene cluster, as described in L. garvieae Lg2 (Miyauchi et al. ), was amplified in isolates A1–A3, A5, A6, A11 and A12 using the TaKaRa LA PCR Kit (TaKaRa Bio Inc. #RR002A, Kyoto, Japan). .. Amplicons were visualised on a 1% agarose gel.

Marker:

Article Title: Loss of the Transcriptional Repressor PAG-3/Gfi-1 Results in Enhanced Neurosecretion that is Dependent on the Dense-Core Vesicle Membrane Protein IDA-1/IA-2
Article Snippet: The gv560 allele was mapped between SNP marker pkP6116 on cosmid F33C8 and pkP6093 on cosmid F01G12. .. Further confirmation of the pag-3(gv560) mutation was provided by rescuing the mutant phenotypes with a 6.5-kb genomic fragment covering the entire coding region of pag-3 that was amplified by long-PCR (TaKaRa LA Taq DNA Polymerase, Takara Bio USA Corporate, Madison, WI).

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    TaKaRa mrna selective pcr kit
    Impact of radial shockwaves on multidifferentiation of MSCs. a Multidifferentiation tests of MSCs showed radial shockwave stimulation increased ALP activity but decreased formation of Oil Red-O-positive lipid droplets, indicating that shockwaves promote osteogenesis induction and block adipogenesis at the same time. No significant differences in the two cohorts of cells after chondrogenic induction. b Impact of shockwaves on MSC differentiation also reflected by <t>mRNA</t> levels of Runx-2 , Osterix , CEBP/α , PPARγ , Sox-9 , and Col-II . Quantitative <t>PCR</t> assay performed at least three times independently; representative result shown. *Statistically significant difference compared with control groups, P
    Mrna Selective Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Impact of radial shockwaves on multidifferentiation of MSCs. a Multidifferentiation tests of MSCs showed radial shockwave stimulation increased ALP activity but decreased formation of Oil Red-O-positive lipid droplets, indicating that shockwaves promote osteogenesis induction and block adipogenesis at the same time. No significant differences in the two cohorts of cells after chondrogenic induction. b Impact of shockwaves on MSC differentiation also reflected by mRNA levels of Runx-2 , Osterix , CEBP/α , PPARγ , Sox-9 , and Col-II . Quantitative PCR assay performed at least three times independently; representative result shown. *Statistically significant difference compared with control groups, P

    Journal: Stem Cell Research & Therapy

    Article Title: Radial shockwave treatment promotes human mesenchymal stem cell self-renewal and enhances cartilage healing

    doi: 10.1186/s13287-018-0805-5

    Figure Lengend Snippet: Impact of radial shockwaves on multidifferentiation of MSCs. a Multidifferentiation tests of MSCs showed radial shockwave stimulation increased ALP activity but decreased formation of Oil Red-O-positive lipid droplets, indicating that shockwaves promote osteogenesis induction and block adipogenesis at the same time. No significant differences in the two cohorts of cells after chondrogenic induction. b Impact of shockwaves on MSC differentiation also reflected by mRNA levels of Runx-2 , Osterix , CEBP/α , PPARγ , Sox-9 , and Col-II . Quantitative PCR assay performed at least three times independently; representative result shown. *Statistically significant difference compared with control groups, P

    Article Snippet: Real-time quantitative PCR analysis Total RNA was extracted from radial-shockwave-treated and nontreated MSCs with TRIzol reagent (Fermentas) and reverse transcribed using the mRNA Selective PCR Kit (TaKaRa).

    Techniques: ALP Assay, Activity Assay, Blocking Assay, Real-time Polymerase Chain Reaction

    Effects of rosuvastatin on electrical remodeling after acute MI. The mRNA expression level of KCND3 potassium ion channel was detected by real-time PCR in atrial tissues following MI, in the sham, MI model, and intervention groups. Compared with the sham

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Effects of rosuvastatin on atrial nerve sprouting and electrical remodeling in rabbits with myocardial infarction

    doi:

    Figure Lengend Snippet: Effects of rosuvastatin on electrical remodeling after acute MI. The mRNA expression level of KCND3 potassium ion channel was detected by real-time PCR in atrial tissues following MI, in the sham, MI model, and intervention groups. Compared with the sham

    Article Snippet: The mRNA expression levels of TH and KCND3were detected with the RT-PCR reagent kit (Takara, Dalian, Liaoning, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Effects of rosuvastatin on TH expression level in atrial tissues after acute MI. The mRNA and protein expression levels of TH in rabbit atrial tissues were detected by real-Time PCR (A) and Western blot analysis (B), respectively. Compared with the sham

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Effects of rosuvastatin on atrial nerve sprouting and electrical remodeling in rabbits with myocardial infarction

    doi:

    Figure Lengend Snippet: Effects of rosuvastatin on TH expression level in atrial tissues after acute MI. The mRNA and protein expression levels of TH in rabbit atrial tissues were detected by real-Time PCR (A) and Western blot analysis (B), respectively. Compared with the sham

    Article Snippet: The mRNA expression levels of TH and KCND3were detected with the RT-PCR reagent kit (Takara, Dalian, Liaoning, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    siRNA (-2752-21) targets tomato SlLNR1 . (A) Schematic illustration of SlLNR1 structure and its associated siRNAs. The sense SlLNR1 , SlLNR1 (+), from the susceptible cultivar is an 1132-nt lncRNA, which is validated by RACE. Its anti-sense, SlLNR1 (-), is generated by sequencing a segmental RT-PCR (the 54 th bp of the 3’ end to the 1008 th ). The full sequence of SlLNR1 was shown in S4 Fig . The vertical bar indicate the target site of siRNA(-2752-21). The data derived from small RNA sequencing of TYLCV infected plants or mock was aligned to SlLNR1 and the number indicates the aligned siRNA reads. (B) Negative correlation of expression of siRNA(-2752-21) and SlLNR1 . SlLNR1 (+) or SlLNR1 (-) was expressed with siRNA(-2752-21) in N . benthamiana . The RNA sample was extracted at 48 h after agroinfiltration. The EF1a gene of N . benthamiana was used as a refernce. (C) SlLNR1 was down regulated in the pTRV2:IR inoculated susceptible plants but not in the resistant plants. The RNA sample was extracted at 15 days after pTRV2:IR and EV inoculated plants. The relative expression of SlLNR1 was measured by qRT-PCR and calculated in relation to EV inoculated plants according to the ΔΔ Ct method. The tomato actin gene was set as reference gene. Error bars represented SE of three biological replicates and significant differences by Student’s t test (*, p

    Journal: PLoS Pathogens

    Article Title: Tomato yellow leaf curl virus intergenic siRNAs target a host long noncoding RNA to modulate disease symptoms

    doi: 10.1371/journal.ppat.1007534

    Figure Lengend Snippet: siRNA (-2752-21) targets tomato SlLNR1 . (A) Schematic illustration of SlLNR1 structure and its associated siRNAs. The sense SlLNR1 , SlLNR1 (+), from the susceptible cultivar is an 1132-nt lncRNA, which is validated by RACE. Its anti-sense, SlLNR1 (-), is generated by sequencing a segmental RT-PCR (the 54 th bp of the 3’ end to the 1008 th ). The full sequence of SlLNR1 was shown in S4 Fig . The vertical bar indicate the target site of siRNA(-2752-21). The data derived from small RNA sequencing of TYLCV infected plants or mock was aligned to SlLNR1 and the number indicates the aligned siRNA reads. (B) Negative correlation of expression of siRNA(-2752-21) and SlLNR1 . SlLNR1 (+) or SlLNR1 (-) was expressed with siRNA(-2752-21) in N . benthamiana . The RNA sample was extracted at 48 h after agroinfiltration. The EF1a gene of N . benthamiana was used as a refernce. (C) SlLNR1 was down regulated in the pTRV2:IR inoculated susceptible plants but not in the resistant plants. The RNA sample was extracted at 15 days after pTRV2:IR and EV inoculated plants. The relative expression of SlLNR1 was measured by qRT-PCR and calculated in relation to EV inoculated plants according to the ΔΔ Ct method. The tomato actin gene was set as reference gene. Error bars represented SE of three biological replicates and significant differences by Student’s t test (*, p

    Article Snippet: The 5’ flanking region of the sense transcripts of SlLNR1 were obtained by RNA ligase-mediated rapid amplification of 5' cDNA ends First Choice ® RLM-RACE Kit (Invitrogen, USA), according to the instructions of the manufacturer, and the 3’ end was verified by 3’ RACE PCR kit (TAKARA).

    Techniques: Generated, Sequencing, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, RNA Sequencing Assay, Infection, Expressing, Quantitative RT-PCR

    Identification of a 25-nt segment and a vsRNA that induce stunt and curled leaves in tomato. (A) VsRNAs generated by the IR and sequence alignment of vsRNAs and SlLNR1 . Location and frequency of TYCLV-derived siRNAs (vsRNAs) were mapped to the IR in sense- (above the x-axis) or antisense- (below the x-axis) orientation. Genome organization of the IR was shown at the top in which the inverted repeat was symbolized as a stem loop. Numbers indicate the first (2616) and last (147) nucleotides of the IR sequence. The histogram of location, frequency and size distribution of vsRNAs corresponding to the 25-nt-fragment (2730–2754) were shown at the medium panel. The fragment was highly complemented with SlLNR1 (-). The scissor means the cleavage site determined by 5’-RACE analysis. (B) Phenotypes of tomato inoculated with pTRV2:4TR. TYLCV-susceptible tomato plants were inoculated with pTRV2 containing 4×25-nt-fragment (2730–2754). The photos were taken at 15 dpi. (C) Validation of siRNA(-2752-21) in the tomato plants by siRNA Northern blot. The leaves of susceptible tomato plants inoculated by TYLCV infectious clone, natural infection by viruliferous whiteflies, agroinfiltrated with pTRV2:4TR and pTRV2:IR were used for total RNA extraction and analyzed at 24 dpi. U6 gene was set as the internal control. (D) Validation of siRNA(-2752-21) presence and downregulation of SlLNR1 in the overexpressed plants. Two individual transgenic lines (pCAMBIA2301:siRNA-1/2) with overexpression of siRNA(-2752-21) were used for total RNA extraction and analyzed. EV indicates the transgenic plant with the EV. The lower panel shows the relative expressi o n of SlLNR1 that was measured by qRT-PCR and calculated in relation to the transgenic plants according to the ΔΔ Ct method using tomat o actin gene as the reference. Error bars represented SE of three biological replicates and significant differences by Student’ s t test (*, p

    Journal: PLoS Pathogens

    Article Title: Tomato yellow leaf curl virus intergenic siRNAs target a host long noncoding RNA to modulate disease symptoms

    doi: 10.1371/journal.ppat.1007534

    Figure Lengend Snippet: Identification of a 25-nt segment and a vsRNA that induce stunt and curled leaves in tomato. (A) VsRNAs generated by the IR and sequence alignment of vsRNAs and SlLNR1 . Location and frequency of TYCLV-derived siRNAs (vsRNAs) were mapped to the IR in sense- (above the x-axis) or antisense- (below the x-axis) orientation. Genome organization of the IR was shown at the top in which the inverted repeat was symbolized as a stem loop. Numbers indicate the first (2616) and last (147) nucleotides of the IR sequence. The histogram of location, frequency and size distribution of vsRNAs corresponding to the 25-nt-fragment (2730–2754) were shown at the medium panel. The fragment was highly complemented with SlLNR1 (-). The scissor means the cleavage site determined by 5’-RACE analysis. (B) Phenotypes of tomato inoculated with pTRV2:4TR. TYLCV-susceptible tomato plants were inoculated with pTRV2 containing 4×25-nt-fragment (2730–2754). The photos were taken at 15 dpi. (C) Validation of siRNA(-2752-21) in the tomato plants by siRNA Northern blot. The leaves of susceptible tomato plants inoculated by TYLCV infectious clone, natural infection by viruliferous whiteflies, agroinfiltrated with pTRV2:4TR and pTRV2:IR were used for total RNA extraction and analyzed at 24 dpi. U6 gene was set as the internal control. (D) Validation of siRNA(-2752-21) presence and downregulation of SlLNR1 in the overexpressed plants. Two individual transgenic lines (pCAMBIA2301:siRNA-1/2) with overexpression of siRNA(-2752-21) were used for total RNA extraction and analyzed. EV indicates the transgenic plant with the EV. The lower panel shows the relative expressi o n of SlLNR1 that was measured by qRT-PCR and calculated in relation to the transgenic plants according to the ΔΔ Ct method using tomat o actin gene as the reference. Error bars represented SE of three biological replicates and significant differences by Student’ s t test (*, p

    Article Snippet: The 5’ flanking region of the sense transcripts of SlLNR1 were obtained by RNA ligase-mediated rapid amplification of 5' cDNA ends First Choice ® RLM-RACE Kit (Invitrogen, USA), according to the instructions of the manufacturer, and the 3’ end was verified by 3’ RACE PCR kit (TAKARA).

    Techniques: Generated, Sequencing, Derivative Assay, Northern Blot, Infection, RNA Extraction, Transgenic Assay, Over Expression, Quantitative RT-PCR

    Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time RT-PCR of Kit mRNA abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition

    Journal: Nature Communications

    Article Title: Sertoli cell-only phenotype and scRNA-seq define PRAMEF12 as a factor essential for spermatogenesis in mice

    doi: 10.1038/s41467-019-13193-3

    Figure Lengend Snippet: Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time RT-PCR of Kit mRNA abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition

    Article Snippet: After linearization by digestion with DraI, the DNA fragment was purified with PCR Clean-up Kit (Clontech Laboratories) and transcribed using AmpliScribe T7-Flash Transcription Kit (Lucigen).

    Techniques: Immunofluorescence, Mouse Assay, Staining, Immunohistochemistry, Quantitative RT-PCR, Two Tailed Test