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TaKaRa pcr reactions
Nucleotide polymorphisms from an alignment of EPSPS cDNA sequences of allohexaploid wheat and wheat progenitors. Vertical lines in the diagram indicate the position of nucleotide polymorphisms identified in a ClustalW multiple sequence alignment of the 1190-bp cDNA clones from T. aestivum ‘Louise’ (T.a_cDNA#), with our 1190-bp cDNA consensus sequences for T. turgidum (AB progenitor) (T.t_cDNA), Ae. tauschii (D progenitor) (Ae.t_cDNA), T. monococcum (A-relative) (T.m_cDNA), and Ae. speltoides (B-relative) (Ae.s_cDNA). The equivalent 1190-bp sequence of the genomic DNA consensus sequences of “TaEPSPS-7A1, TaEPSPS-7D1, and TaEPSPS-4A1” are shown with intron sequences removed. Only the TaEPSPS-7A1 and TaEPSPS-7D1 cDNAs were recovered by <t>F3-R1</t> <t>PCR</t> amplification. The corresponding 1190-bp TaEPSPS-4A1 cDNA sequence (T.a_cDNA13) was derived from an independent experiment
Pcr Reactions, supplied by TaKaRa, used in various techniques. Bioz Stars score: 95/100, based on 30259 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Molecular and phylogenetic characterization of the homoeologous EPSP Synthase genes of allohexaploid wheat, Triticum aestivum (L.)"

Article Title: Molecular and phylogenetic characterization of the homoeologous EPSP Synthase genes of allohexaploid wheat, Triticum aestivum (L.)

Journal: BMC Genomics

doi: 10.1186/s12864-015-2084-1

Nucleotide polymorphisms from an alignment of EPSPS cDNA sequences of allohexaploid wheat and wheat progenitors. Vertical lines in the diagram indicate the position of nucleotide polymorphisms identified in a ClustalW multiple sequence alignment of the 1190-bp cDNA clones from T. aestivum ‘Louise’ (T.a_cDNA#), with our 1190-bp cDNA consensus sequences for T. turgidum (AB progenitor) (T.t_cDNA), Ae. tauschii (D progenitor) (Ae.t_cDNA), T. monococcum (A-relative) (T.m_cDNA), and Ae. speltoides (B-relative) (Ae.s_cDNA). The equivalent 1190-bp sequence of the genomic DNA consensus sequences of “TaEPSPS-7A1, TaEPSPS-7D1, and TaEPSPS-4A1” are shown with intron sequences removed. Only the TaEPSPS-7A1 and TaEPSPS-7D1 cDNAs were recovered by F3-R1 PCR amplification. The corresponding 1190-bp TaEPSPS-4A1 cDNA sequence (T.a_cDNA13) was derived from an independent experiment
Figure Legend Snippet: Nucleotide polymorphisms from an alignment of EPSPS cDNA sequences of allohexaploid wheat and wheat progenitors. Vertical lines in the diagram indicate the position of nucleotide polymorphisms identified in a ClustalW multiple sequence alignment of the 1190-bp cDNA clones from T. aestivum ‘Louise’ (T.a_cDNA#), with our 1190-bp cDNA consensus sequences for T. turgidum (AB progenitor) (T.t_cDNA), Ae. tauschii (D progenitor) (Ae.t_cDNA), T. monococcum (A-relative) (T.m_cDNA), and Ae. speltoides (B-relative) (Ae.s_cDNA). The equivalent 1190-bp sequence of the genomic DNA consensus sequences of “TaEPSPS-7A1, TaEPSPS-7D1, and TaEPSPS-4A1” are shown with intron sequences removed. Only the TaEPSPS-7A1 and TaEPSPS-7D1 cDNAs were recovered by F3-R1 PCR amplification. The corresponding 1190-bp TaEPSPS-4A1 cDNA sequence (T.a_cDNA13) was derived from an independent experiment

Techniques Used: Sequencing, Clone Assay, Polymerase Chain Reaction, Amplification, Derivative Assay

Optimization of the GC-rich TaEPSPS-7A1 PCR amplification. Louise genomic DNA was amplified using the F1.2-R1 primer pair with the indicated buffers designed for amplification of difficult templates (GCI, GCII, D, E, F, G, H, and I), and the indicated concentrations of DMSO (0 %, 2.5 %, 5 %, and 7.5 %). ‘M’ stands for the 1 kb DNA ladder (Thermo Scientific)
Figure Legend Snippet: Optimization of the GC-rich TaEPSPS-7A1 PCR amplification. Louise genomic DNA was amplified using the F1.2-R1 primer pair with the indicated buffers designed for amplification of difficult templates (GCI, GCII, D, E, F, G, H, and I), and the indicated concentrations of DMSO (0 %, 2.5 %, 5 %, and 7.5 %). ‘M’ stands for the 1 kb DNA ladder (Thermo Scientific)

Techniques Used: Polymerase Chain Reaction, Amplification

2) Product Images from "Establishment of nDart1-tagged lines of Koshihikari, an elite variety of rice in Japan"

Article Title: Establishment of nDart1-tagged lines of Koshihikari, an elite variety of rice in Japan

Journal: Breeding Science

doi: 10.1270/jsbbs.19049

Confirmation of nDart1-0 insertion revealed by TAIL-PCR. (A) GS (Green sector) and WS (White sector) in albino of Fig. 2B . (B) Banding pattern of WT, GS and WS using specific primers for each clone. White arrow and arrowhead indicate amplicons with nDart1 insertion and without nDart1 , respectively.
Figure Legend Snippet: Confirmation of nDart1-0 insertion revealed by TAIL-PCR. (A) GS (Green sector) and WS (White sector) in albino of Fig. 2B . (B) Banding pattern of WT, GS and WS using specific primers for each clone. White arrow and arrowhead indicate amplicons with nDart1 insertion and without nDart1 , respectively.

Techniques Used: Polymerase Chain Reaction

3) Product Images from "Establishment of nDart1-tagged lines of Koshihikari, an elite variety of rice in Japan"

Article Title: Establishment of nDart1-tagged lines of Koshihikari, an elite variety of rice in Japan

Journal: Breeding Science

doi: 10.1270/jsbbs.19049

Confirmation of nDart1-0 insertion revealed by TAIL-PCR. (A) GS (Green sector) and WS (White sector) in albino of Fig. 2B . (B) Banding pattern of WT, GS and WS using specific primers for each clone. White arrow and arrowhead indicate amplicons with nDart1 insertion and without nDart1 , respectively.
Figure Legend Snippet: Confirmation of nDart1-0 insertion revealed by TAIL-PCR. (A) GS (Green sector) and WS (White sector) in albino of Fig. 2B . (B) Banding pattern of WT, GS and WS using specific primers for each clone. White arrow and arrowhead indicate amplicons with nDart1 insertion and without nDart1 , respectively.

Techniques Used: Polymerase Chain Reaction

4) Product Images from "Molecular and phylogenetic characterization of the homoeologous EPSP Synthase genes of allohexaploid wheat, Triticum aestivum (L.)"

Article Title: Molecular and phylogenetic characterization of the homoeologous EPSP Synthase genes of allohexaploid wheat, Triticum aestivum (L.)

Journal: BMC Genomics

doi: 10.1186/s12864-015-2084-1

Exon-intron structure of the wheat EPSPS genes compared to OsEPSPS . The boxes and solid lines represent exons (E#) and introns (I#), respectively. Numbers indicate exon and intron size in bp. The cleavage site of the chloroplastic transit signal peptide (cTP) predicted by PredSL is shown. The positions of the primers F1.2, F3, and R1 (arrows) used for initial genomic and cDNA cloning are labeled
Figure Legend Snippet: Exon-intron structure of the wheat EPSPS genes compared to OsEPSPS . The boxes and solid lines represent exons (E#) and introns (I#), respectively. Numbers indicate exon and intron size in bp. The cleavage site of the chloroplastic transit signal peptide (cTP) predicted by PredSL is shown. The positions of the primers F1.2, F3, and R1 (arrows) used for initial genomic and cDNA cloning are labeled

Techniques Used: Clone Assay, Labeling

Optimization of the GC-rich TaEPSPS-7A1 PCR amplification. Louise genomic DNA was amplified using the F1.2-R1 primer pair with the indicated buffers designed for amplification of difficult templates (GCI, GCII, D, E, F, G, H, and I), and the indicated concentrations of DMSO (0 %, 2.5 %, 5 %, and 7.5 %). ‘M’ stands for the 1 kb DNA ladder (Thermo Scientific)
Figure Legend Snippet: Optimization of the GC-rich TaEPSPS-7A1 PCR amplification. Louise genomic DNA was amplified using the F1.2-R1 primer pair with the indicated buffers designed for amplification of difficult templates (GCI, GCII, D, E, F, G, H, and I), and the indicated concentrations of DMSO (0 %, 2.5 %, 5 %, and 7.5 %). ‘M’ stands for the 1 kb DNA ladder (Thermo Scientific)

Techniques Used: Polymerase Chain Reaction, Amplification

Related Articles

Amplification:

Article Title: Essential Domains for Ribonucleoprotein Complex Formation Required for Retrotransposition of Telomere-Specific Non-Long Terminal Repeat Retrotransposon SART1 †
Article Snippet: .. After heat inactivation at 80°C for 20 min, 1 μl of the reaction mixture was amplified by PCR for 35 cycles of 98°C for 20 s, 60°C for 30 s, and 72°C for 30 s, with an initial denaturation at 96°C for 2 min and a final extension at 72°C for 5 min. PCR mixtures contained 200 μM concentrations of each dNTP, 2 mM MgCl2 , 50 mM KCl, 0.5 U of Ex- Taq (TaKaRa), and 0.5 μM concentrations of each primer (see Table S1 in the supplemental material). .. Purified SART1 complex was overlaid onto 3.3 ml of 10 to 40% glycerol gradient in the HG500 buffer.

Article Title: Association between polymorphisms in the GRIN1 gene 5′ regulatory region and schizophrenia in a northern Han Chinese population and haplotype effects on protein expression in vitro
Article Snippet: .. Genomic DNA (1 μL, about 30 ng) was amplified under the following reaction contents: 1 μL (5 pmol) each of sense and antisense primers, 2 μL (3 nmol) of dNTP mix, 0.2 μL (about 0.5 U) of PrimeSTAR® HS DNA polymerase (Takara, Dalian, China) and 10 μL 2 × Prime STAR HS GC buffer. ..

DNA Synthesis:

Article Title: Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease
Article Snippet: .. Resumption of DNA Synthesis by DNA Polymerase after the Removal of CTNAs The hybridized oligonucleotides (400 fmol) were first treated with ERCC1-XPF (230 fmol) in 10 µL of 50 mM Tris-HCl buffer (pH 8.0), containing 2 mM MgCl2 , 0.5 mM DTT and 0.1 mg·mL−1 BSA, at 25 °C for 16 h. To the reaction mixture was added 5 µL of 30 mM Tris-HCl buffer (pH 7.9), containing 150 mM NaCl, 30 mM MgCl2 , 30 mM DTT, 300 µM dNTPs and the Klenow fragment of Escherichia coli DNA polymerase I, lacking the 3′–5′ exonuclease activity (KF− , 0.1 unit, Takara Bio). ..

Marker:

Article Title: Small Molecular Contaminant and Microorganism Can Be Simultaneously Detected Based on Nanobody-Phage: Using Carcinogen Aflatoxin and Its Main Fungal Aspergillus Section Flavi spp. in Stored Maize for Demonstration
Article Snippet: .. DNA polymerase (iTaq), Mg2+ , dNTPs, 6× loading buffer, and DNA marker were bought from Takara Bio (Beijing, China). .. The anti-aflatoxins monoclonal antibody 1C11 (mAb 1C11) and V2–5 phage displaying nanobody specific for 1C11 were produced by our team ( ; ).

Activity Assay:

Article Title: Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease
Article Snippet: .. Resumption of DNA Synthesis by DNA Polymerase after the Removal of CTNAs The hybridized oligonucleotides (400 fmol) were first treated with ERCC1-XPF (230 fmol) in 10 µL of 50 mM Tris-HCl buffer (pH 8.0), containing 2 mM MgCl2 , 0.5 mM DTT and 0.1 mg·mL−1 BSA, at 25 °C for 16 h. To the reaction mixture was added 5 µL of 30 mM Tris-HCl buffer (pH 7.9), containing 150 mM NaCl, 30 mM MgCl2 , 30 mM DTT, 300 µM dNTPs and the Klenow fragment of Escherichia coli DNA polymerase I, lacking the 3′–5′ exonuclease activity (KF− , 0.1 unit, Takara Bio). ..

Polymerase Chain Reaction:

Article Title: Essential Domains for Ribonucleoprotein Complex Formation Required for Retrotransposition of Telomere-Specific Non-Long Terminal Repeat Retrotransposon SART1 †
Article Snippet: .. After heat inactivation at 80°C for 20 min, 1 μl of the reaction mixture was amplified by PCR for 35 cycles of 98°C for 20 s, 60°C for 30 s, and 72°C for 30 s, with an initial denaturation at 96°C for 2 min and a final extension at 72°C for 5 min. PCR mixtures contained 200 μM concentrations of each dNTP, 2 mM MgCl2 , 50 mM KCl, 0.5 U of Ex- Taq (TaKaRa), and 0.5 μM concentrations of each primer (see Table S1 in the supplemental material). .. Purified SART1 complex was overlaid onto 3.3 ml of 10 to 40% glycerol gradient in the HG500 buffer.

Article Title: Functional variations of the TLR4 gene in association with chronic obstructive pulmonary disease and pulmonary tuberculosis
Article Snippet: .. PCR was performed to amplify the fragments of Wt, Mut1 and Mut2 in 0.5 ml tubes that included 0.5 μl of PrimeSTAR HS DNA polymerase (Takara), 1 μl of 10 μM forward and reverse primers, 4 μl of 2.5 mM each dNTP Mix (Takara), 10 ng of genomic DNA and 10 μl of 5 × PS Buffer (Takara). ..

Plasmid Preparation:

Article Title: A transcription and translation-coupled DNA replication system using rolling-circle replication
Article Snippet: .. Assay of the TTcDR reaction The optimized composition of the TTcDR system was as follows: template plasmid DNA (1 ng/μl), dNTPs (0.3 mM each, Takara), [32 -P] dCTP (3.3 μM, PerkinElmer), magnesium acetate (7.9 mM, Wako), potassium glutamate (70 mM, Wako), spermidine (0.375 mM, Nakarai), dithiothreitol (6 mM, Nakarai), ATP (0.375 mM, GE Healthcare), GTP (0.25 mM, GE Healthcare), CTP (0.125 mM, GE Healthcare), UTP (0.125 mM, GE Healthcare), creatine phosphate (25 mM, Nakarai), E. coli tRNA mixture (0.518 μg/μl, Roche), 10-formyl-5,6,7,8-tetrahydrofolic acid (10 ng/μl), Cys (0.3 mM, Wako), Tyr (0.3 mM, Wako), the other 18 amino acids except for Cys and Tyr (0.36 mM, Wako), 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (100 mM, pH 7.6, Sigma), ribosomes (1 μM), IF1 (25 μM), IF2 (1 μM), IF3 (4.9 μM), EF-G (1.1 μM), EF-Tu (80 μM), EF-Ts (3.3 μM), RF1 (0.05 μM), RF2 (0.05 μM), RF3 (0.17 μM), RRF (3.9 μM), AlaRS (730 nM), ArgRS (30 nM), AsnRS (420 nM), AspRS (120 nM), CysRS (20 nM), GlnRS (60 nM), GluRS (230 nM), GlyRS (90 nM), HisRS (90 nM), IleRS (370 nM), LeuRS (40 nM), LysRS (120 nM), MetRS (110 nM), PheRS (130 nM), ProRS (170 nM), SerRS (80 nM), ThrRS (80 nM), TrpRS (30 nM), TyrRS (150 nM), ValRS (20 nM), MTF (590 nM), creatine kinase (0.25 μM), myokinase (1.4 μM), nucleoside-diphosphate kinase (20 nM), pyrophosphatase (40 nM), yeast inorganic pyrophosphatase (0.2 mU/μl, New England BioLabs (NEB)), ribonuclease inhibitor (0.1 U/μl; Promega), and T7 RNA polymerase (0.42 U/μl; Takara). .. Most of the proteins described above were purified according to a previously described method except for yeast inorganic pyrophosphatase, ribonuclease inhibitor, and T7 RNA polymerase.

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