long range pcr kit  (TaKaRa)

 
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    Name:
    LA PCR Kit
    Description:
    The LA PCR Kit Version 2 1 contains all the reagents needed for amplification of long DNA templates enabling routine amplification of products up to 20 kb in length including GC rich amplicons For some DNA templates amplification of up to 48 kb is possible This long range PCR kit contains TaKaRa LA Taq DNA Polymerase buffers MgCl2 dNTPs molecular weight markers and control templates plus corresponding primers to ensure optimal PCR performance during long PCR
    Catalog Number:
    rr013b
    Price:
    None
    Size:
    100 Rxns
    Category:
    LA Taq PCR kit LA Taq products Long range PCR PCR
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    Structured Review

    TaKaRa long range pcr kit
    The LA PCR Kit Version 2 1 contains all the reagents needed for amplification of long DNA templates enabling routine amplification of products up to 20 kb in length including GC rich amplicons For some DNA templates amplification of up to 48 kb is possible This long range PCR kit contains TaKaRa LA Taq DNA Polymerase buffers MgCl2 dNTPs molecular weight markers and control templates plus corresponding primers to ensure optimal PCR performance during long PCR
    https://www.bioz.com/result/long range pcr kit/product/TaKaRa
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    long range pcr kit - by Bioz Stars, 2020-08
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro
    Article Snippet: .. Single-stranded cDNA was synthesized by reverse transcriptase (RT) with random hexamer oligonucleotides as primers, and the desired cDNA was amplified by polymerase chain reaction (PCR) using a TaKaRa RNA LA PCR kit (TaKaRa Biomedicals, Kyoto, Japan). .. Primers (sense and antisense) for the amplification of each cytokine cDNA were as follows: β-actin sense, TGG AAT CCT GTG GCA TCC ATG AAA C; β-actin antisense, TAA AAC GCA GCT CAG TAA CAG TCC G; IL-1α sense, CTC TAG AGC ACC ATG CTA CAG AC; IL-1α antisense, TGG AAT CCA GGG GAA ACA CTG; IL-2 sense, TGA TGG ACC TAC AGG AGC TCC TGA G; IL-2 antisense, GAG TCA AAT CCA GAA CAT GCC GCA G; IL-3 sense, GAA GTG GAT CCT GAG GAC AGA TAC G; IL-3 antisense, GAC CAT GGG CCA TGA GGA ACA TTC; IL-4 sense, CGA AGA ACA CCA CAG AGA GTG AGC T; IL-4 antisense, GAC TCA TTC ATG GTG CAG CTT ATC G; IL-5 sense, CTC TAG TAA GCC CAC TTC TA; IL-5 antisense, TGA TAC ATG AAT AAC ATC CC; IL-6 sense, TGG AGT CAC AGA AGG AGT GGC TAA G; IL-6 antisense, TCT GAC CAC AGA AGG AGT GGC TAA G; IL-7 sense, AAA TGC AGC TGA CTG CTG GC; IL-7 antisense, TCT CCA GTC TAA AAC AGG AC; IL-10 sense, TAC CTG GTA GAA GTG ATG CC; IL-10 antisense, CAT CAT GTA TGC TTC TAT GC; tumour necrosis factor-α (TNF-α) sense, GGC AGG TCT ACT TTG GAG TCA TTG C; TNF-α antisense, ACA TTC GAG GCT CCA GTG AAT TCG G; lymphotoxin (LT)-α sense, TGG CTG GGA ACA GGG GAA GGT TGA C; LT-α antisense, CGT GCT TTC TTC TAG AAC CCC TTG G. The conditions for PCR were 1 min at 94°, 2 min at 55° and 3 min at 72° for 30 cycles.

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells
    Article Snippet: .. RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume. .. The gene of interest was amplified by ExTaq polymerase in PCR kits using primer sequences described in the Additional file .

    Article Title: Tomato yellow leaf curl virus intergenic siRNAs target a host long noncoding RNA to modulate disease symptoms
    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). .. Full sense transcript of the SlLNR1 was amplified with the primers designed according to the joint sequence by RT-PCR.

    Article Title: Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation
    Article Snippet: .. The sequence of the gene coding the gametocyte protein was amplified by reverse transcription polymerase chain reaction (RT-PCR) using the RNA LA PCR Kit (TaKaRa Bio. ..

    Synthesized:

    Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro
    Article Snippet: .. Single-stranded cDNA was synthesized by reverse transcriptase (RT) with random hexamer oligonucleotides as primers, and the desired cDNA was amplified by polymerase chain reaction (PCR) using a TaKaRa RNA LA PCR kit (TaKaRa Biomedicals, Kyoto, Japan). .. Primers (sense and antisense) for the amplification of each cytokine cDNA were as follows: β-actin sense, TGG AAT CCT GTG GCA TCC ATG AAA C; β-actin antisense, TAA AAC GCA GCT CAG TAA CAG TCC G; IL-1α sense, CTC TAG AGC ACC ATG CTA CAG AC; IL-1α antisense, TGG AAT CCA GGG GAA ACA CTG; IL-2 sense, TGA TGG ACC TAC AGG AGC TCC TGA G; IL-2 antisense, GAG TCA AAT CCA GAA CAT GCC GCA G; IL-3 sense, GAA GTG GAT CCT GAG GAC AGA TAC G; IL-3 antisense, GAC CAT GGG CCA TGA GGA ACA TTC; IL-4 sense, CGA AGA ACA CCA CAG AGA GTG AGC T; IL-4 antisense, GAC TCA TTC ATG GTG CAG CTT ATC G; IL-5 sense, CTC TAG TAA GCC CAC TTC TA; IL-5 antisense, TGA TAC ATG AAT AAC ATC CC; IL-6 sense, TGG AGT CAC AGA AGG AGT GGC TAA G; IL-6 antisense, TCT GAC CAC AGA AGG AGT GGC TAA G; IL-7 sense, AAA TGC AGC TGA CTG CTG GC; IL-7 antisense, TCT CCA GTC TAA AAC AGG AC; IL-10 sense, TAC CTG GTA GAA GTG ATG CC; IL-10 antisense, CAT CAT GTA TGC TTC TAT GC; tumour necrosis factor-α (TNF-α) sense, GGC AGG TCT ACT TTG GAG TCA TTG C; TNF-α antisense, ACA TTC GAG GCT CCA GTG AAT TCG G; lymphotoxin (LT)-α sense, TGG CTG GGA ACA GGG GAA GGT TGA C; LT-α antisense, CGT GCT TTC TTC TAG AAC CCC TTG G. The conditions for PCR were 1 min at 94°, 2 min at 55° and 3 min at 72° for 30 cycles.

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells
    Article Snippet: .. RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume. .. The gene of interest was amplified by ExTaq polymerase in PCR kits using primer sequences described in the Additional file .

    Isolation:

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells
    Article Snippet: .. RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume. .. The gene of interest was amplified by ExTaq polymerase in PCR kits using primer sequences described in the Additional file .

    Produced:

    Article Title: The Bmdsx transgene including trimmed introns is sex-specifically spliced in tissues of the silkworm, Bombyx mori
    Article Snippet: .. RT-PCR was performed using the LA RNA PCR kit (Takara, www.takara-bio.co.jp ) following the manufacturer's instructions. cDNA was produced by random priming. .. RT-PCR primers were as follows: endogenous BmA3 , BmA3QPCR1F (5′-TACAATGAGCTGCGTGTCG-3′) and BmA3QPCR1R (5′-CGGGCGTGTTGAATGTTTC -3′); and Bmdsx mRNA transcribed from the transgene, TGM2F (5′-ATTGGCGGGACACGATC-3′) and TGM2R (5′-AGCGCTCCGTAGCACAA-3′).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells
    Article Snippet: .. RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume. .. The gene of interest was amplified by ExTaq polymerase in PCR kits using primer sequences described in the Additional file .

    Article Title: The Bmdsx transgene including trimmed introns is sex-specifically spliced in tissues of the silkworm, Bombyx mori
    Article Snippet: .. RT-PCR was performed using the LA RNA PCR kit (Takara, www.takara-bio.co.jp ) following the manufacturer's instructions. cDNA was produced by random priming. .. RT-PCR primers were as follows: endogenous BmA3 , BmA3QPCR1F (5′-TACAATGAGCTGCGTGTCG-3′) and BmA3QPCR1R (5′-CGGGCGTGTTGAATGTTTC -3′); and Bmdsx mRNA transcribed from the transgene, TGM2F (5′-ATTGGCGGGACACGATC-3′) and TGM2R (5′-AGCGCTCCGTAGCACAA-3′).

    Article Title: Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation
    Article Snippet: .. The sequence of the gene coding the gametocyte protein was amplified by reverse transcription polymerase chain reaction (RT-PCR) using the RNA LA PCR Kit (TaKaRa Bio. ..

    Article Title: Spermidine Synthase Genes Are Essential for Survival of Arabidopsis
    Article Snippet: .. RT-PCR was conducted by using the RNA LA PCR Kit (Takara, Kyoto) with 0.5 μ g of total RNA. ..

    Random Hexamer Labeling:

    Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro
    Article Snippet: .. Single-stranded cDNA was synthesized by reverse transcriptase (RT) with random hexamer oligonucleotides as primers, and the desired cDNA was amplified by polymerase chain reaction (PCR) using a TaKaRa RNA LA PCR kit (TaKaRa Biomedicals, Kyoto, Japan). .. Primers (sense and antisense) for the amplification of each cytokine cDNA were as follows: β-actin sense, TGG AAT CCT GTG GCA TCC ATG AAA C; β-actin antisense, TAA AAC GCA GCT CAG TAA CAG TCC G; IL-1α sense, CTC TAG AGC ACC ATG CTA CAG AC; IL-1α antisense, TGG AAT CCA GGG GAA ACA CTG; IL-2 sense, TGA TGG ACC TAC AGG AGC TCC TGA G; IL-2 antisense, GAG TCA AAT CCA GAA CAT GCC GCA G; IL-3 sense, GAA GTG GAT CCT GAG GAC AGA TAC G; IL-3 antisense, GAC CAT GGG CCA TGA GGA ACA TTC; IL-4 sense, CGA AGA ACA CCA CAG AGA GTG AGC T; IL-4 antisense, GAC TCA TTC ATG GTG CAG CTT ATC G; IL-5 sense, CTC TAG TAA GCC CAC TTC TA; IL-5 antisense, TGA TAC ATG AAT AAC ATC CC; IL-6 sense, TGG AGT CAC AGA AGG AGT GGC TAA G; IL-6 antisense, TCT GAC CAC AGA AGG AGT GGC TAA G; IL-7 sense, AAA TGC AGC TGA CTG CTG GC; IL-7 antisense, TCT CCA GTC TAA AAC AGG AC; IL-10 sense, TAC CTG GTA GAA GTG ATG CC; IL-10 antisense, CAT CAT GTA TGC TTC TAT GC; tumour necrosis factor-α (TNF-α) sense, GGC AGG TCT ACT TTG GAG TCA TTG C; TNF-α antisense, ACA TTC GAG GCT CCA GTG AAT TCG G; lymphotoxin (LT)-α sense, TGG CTG GGA ACA GGG GAA GGT TGA C; LT-α antisense, CGT GCT TTC TTC TAG AAC CCC TTG G. The conditions for PCR were 1 min at 94°, 2 min at 55° and 3 min at 72° for 30 cycles.

    Polymerase Chain Reaction:

    Article Title: Murine lymph node-derived stromal cells effectively support survival but induce no activation/proliferation of peripheral resting T cells in vitro
    Article Snippet: .. Single-stranded cDNA was synthesized by reverse transcriptase (RT) with random hexamer oligonucleotides as primers, and the desired cDNA was amplified by polymerase chain reaction (PCR) using a TaKaRa RNA LA PCR kit (TaKaRa Biomedicals, Kyoto, Japan). .. Primers (sense and antisense) for the amplification of each cytokine cDNA were as follows: β-actin sense, TGG AAT CCT GTG GCA TCC ATG AAA C; β-actin antisense, TAA AAC GCA GCT CAG TAA CAG TCC G; IL-1α sense, CTC TAG AGC ACC ATG CTA CAG AC; IL-1α antisense, TGG AAT CCA GGG GAA ACA CTG; IL-2 sense, TGA TGG ACC TAC AGG AGC TCC TGA G; IL-2 antisense, GAG TCA AAT CCA GAA CAT GCC GCA G; IL-3 sense, GAA GTG GAT CCT GAG GAC AGA TAC G; IL-3 antisense, GAC CAT GGG CCA TGA GGA ACA TTC; IL-4 sense, CGA AGA ACA CCA CAG AGA GTG AGC T; IL-4 antisense, GAC TCA TTC ATG GTG CAG CTT ATC G; IL-5 sense, CTC TAG TAA GCC CAC TTC TA; IL-5 antisense, TGA TAC ATG AAT AAC ATC CC; IL-6 sense, TGG AGT CAC AGA AGG AGT GGC TAA G; IL-6 antisense, TCT GAC CAC AGA AGG AGT GGC TAA G; IL-7 sense, AAA TGC AGC TGA CTG CTG GC; IL-7 antisense, TCT CCA GTC TAA AAC AGG AC; IL-10 sense, TAC CTG GTA GAA GTG ATG CC; IL-10 antisense, CAT CAT GTA TGC TTC TAT GC; tumour necrosis factor-α (TNF-α) sense, GGC AGG TCT ACT TTG GAG TCA TTG C; TNF-α antisense, ACA TTC GAG GCT CCA GTG AAT TCG G; lymphotoxin (LT)-α sense, TGG CTG GGA ACA GGG GAA GGT TGA C; LT-α antisense, CGT GCT TTC TTC TAG AAC CCC TTG G. The conditions for PCR were 1 min at 94°, 2 min at 55° and 3 min at 72° for 30 cycles.

    Article Title: B-cell translocation gene 2 mediates crosstalk between PI3K/Akt1 and NF?B pathways which enhances transcription of MnSOD by accelerating I?B? degradation in normal and cancer cells
    Article Snippet: .. RT-PCR Total cellular RNAs (1.0 μg) isolated with RNAiso Plus were used for cDNA preparation and then amplified by PCR kit (Takara Inc., Japan); First strand cDNA was synthesized using oligo-dT by reverse transcription reaction in 10 μl of reaction volume. .. The gene of interest was amplified by ExTaq polymerase in PCR kits using primer sequences described in the Additional file .

    Article Title: The Riemerella anatipestifer M949_RS01035 gene is involved in bacterial lipopolysaccharide biosynthesis
    Article Snippet: .. Primer pairs specific for Tn4351 (primers TN-1 and IS4351-F) were used to amplify the sequences adjacent to the insertion site using the LA PCR kit (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China). .. The nucleotide sequence was compared to sequence in the National Center for Biotechnology Information database using the BLASTX program [ ].

    Article Title: The Bmdsx transgene including trimmed introns is sex-specifically spliced in tissues of the silkworm, Bombyx mori
    Article Snippet: .. RT-PCR was performed using the LA RNA PCR kit (Takara, www.takara-bio.co.jp ) following the manufacturer's instructions. cDNA was produced by random priming. .. RT-PCR primers were as follows: endogenous BmA3 , BmA3QPCR1F (5′-TACAATGAGCTGCGTGTCG-3′) and BmA3QPCR1R (5′-CGGGCGTGTTGAATGTTTC -3′); and Bmdsx mRNA transcribed from the transgene, TGM2F (5′-ATTGGCGGGACACGATC-3′) and TGM2R (5′-AGCGCTCCGTAGCACAA-3′).

    Article Title: Tomato yellow leaf curl virus intergenic siRNAs target a host long noncoding RNA to modulate disease symptoms
    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). .. Full sense transcript of the SlLNR1 was amplified with the primers designed according to the joint sequence by RT-PCR.

    Article Title: Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation
    Article Snippet: .. The sequence of the gene coding the gametocyte protein was amplified by reverse transcription polymerase chain reaction (RT-PCR) using the RNA LA PCR Kit (TaKaRa Bio. ..

    Article Title: Spermidine Synthase Genes Are Essential for Survival of Arabidopsis
    Article Snippet: .. RT-PCR was conducted by using the RNA LA PCR Kit (Takara, Kyoto) with 0.5 μ g of total RNA. ..

    Article Title: Characterization of a novel germline PALB2 duplication in a hereditary breast and ovarian cancer family
    Article Snippet: .. LR-PCR was performed using a forward primer in intron 12 and a reverse primer in 3′-untranslated region (UTR) of PALB2 and the TaKaRa LA PCR kit (TaKaRa, Clontech) following the manufacturer’s instructions. ..

    Sequencing:

    Article Title: Cloning and characterization of an Eimeria necatrix gene encoding a gametocyte protein and associated with oocyst wall formation
    Article Snippet: .. The sequence of the gene coding the gametocyte protein was amplified by reverse transcription polymerase chain reaction (RT-PCR) using the RNA LA PCR Kit (TaKaRa Bio. ..

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  • 99
    TaKaRa race pcr kit
    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 <t>RACE.</t> Its anti-sense, SlLNR1 (-), is generated by sequencing a segmental <t>RT-PCR</t> (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
    Race Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/race pcr kit/product/TaKaRa
    Average 99 stars, based on 56 article reviews
    Price from $9.99 to $1999.99
    race pcr kit - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    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

    Identification of the mutant strain RA1062. A PCR amplification. M: Takara DL2000 marker; lanes 1–2: R. anatipestifer 16S rRNA was amplified from the WT strain CH3 (lane 1), the mutant strain RA1062 (lane 2), showing a 744-bp fragment of 16S rRNA; lanes 4–5: a 678-bp fragment of M949_RS01035 was amplified from the WT strain CH3 (lane 4), but not the mutant strain RA1062 (lane 5); lanes 7–8: the 644-bp fragment of erm gene was not amplified from the WT strain CH3 (lane 7), but amplified from the mutant strain RA1062 (lane 8); lanes 3, 6 and 9: the avian pathogenic E. coli strain (APEC, CVCC1547), as negative controls. B Southern blot analysis of the transposon Tn4351 insertion. Lane 1, 10 μg of pEP 4351 digested with Xba I (positive control); Lane 2, 10 μg of chromosomal DNA from mutant strain RA1062 digested with Xba I; Lane 3, 10 μg of chromosomal DNA from the WT strain CH3 digested with Xba I (negative control). The digested sample was resolved on a 0.7% agarose gel and Southern blot analysis was performed using a TnDIG-labeled probe. C Schematic chart of Tn4351 insertion in RA1062 chromosome at 318 bp of the gene, which is 678 nucleotides in length. D qPCR analysis. The expression of the mRNAs were expressed as fold change and calculated using the comparative C T (2 −∆∆CT ) method. Data were normalized to the housekeeping gene ldh and expressed as fold changes. The expression of M949_RS01035 in the mutant strain RA1062 was disrupted. However, no change was shown for its upstream M949_RS10475 gene and downstream M949_RS01030 gene. Error bars represent standard deviations from three replicates (*** p

    Journal: Veterinary Research

    Article Title: The Riemerella anatipestifer M949_RS01035 gene is involved in bacterial lipopolysaccharide biosynthesis

    doi: 10.1186/s13567-018-0589-8

    Figure Lengend Snippet: Identification of the mutant strain RA1062. A PCR amplification. M: Takara DL2000 marker; lanes 1–2: R. anatipestifer 16S rRNA was amplified from the WT strain CH3 (lane 1), the mutant strain RA1062 (lane 2), showing a 744-bp fragment of 16S rRNA; lanes 4–5: a 678-bp fragment of M949_RS01035 was amplified from the WT strain CH3 (lane 4), but not the mutant strain RA1062 (lane 5); lanes 7–8: the 644-bp fragment of erm gene was not amplified from the WT strain CH3 (lane 7), but amplified from the mutant strain RA1062 (lane 8); lanes 3, 6 and 9: the avian pathogenic E. coli strain (APEC, CVCC1547), as negative controls. B Southern blot analysis of the transposon Tn4351 insertion. Lane 1, 10 μg of pEP 4351 digested with Xba I (positive control); Lane 2, 10 μg of chromosomal DNA from mutant strain RA1062 digested with Xba I; Lane 3, 10 μg of chromosomal DNA from the WT strain CH3 digested with Xba I (negative control). The digested sample was resolved on a 0.7% agarose gel and Southern blot analysis was performed using a TnDIG-labeled probe. C Schematic chart of Tn4351 insertion in RA1062 chromosome at 318 bp of the gene, which is 678 nucleotides in length. D qPCR analysis. The expression of the mRNAs were expressed as fold change and calculated using the comparative C T (2 −∆∆CT ) method. Data were normalized to the housekeeping gene ldh and expressed as fold changes. The expression of M949_RS01035 in the mutant strain RA1062 was disrupted. However, no change was shown for its upstream M949_RS10475 gene and downstream M949_RS01030 gene. Error bars represent standard deviations from three replicates (*** p

    Article Snippet: Primer pairs specific for Tn4351 (primers TN-1 and IS4351-F) were used to amplify the sequences adjacent to the insertion site using the LA PCR kit (TaKaRa Biotechnology (Dalian) Co., Ltd., Dalian, China).

    Techniques: Mutagenesis, Polymerase Chain Reaction, Amplification, Marker, Southern Blot, Positive Control, Negative Control, Agarose Gel Electrophoresis, Labeling, Real-time Polymerase Chain Reaction, Expressing

    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

    Pre-mRNA transcribed from the Bmdsx mini gene is sex-specifically spliced. (A) The diagram shows the structure of the Bmdsx minigene. Open boxes: common exons. Shaded boxes: female-specific exons. Arrows: primers for PCR. (B) Poly (A)+RNA was extracted from the fat body of transgenic silkworms. These RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers TGM2F and TGM2R. Resulting products were separated on a 1% agarose gel and visualized with SYBR Green I (Molecular Probes) at a dilution of 1: 10,000. M represents the DNA marker(λ /Hind III+ φX174/ Hinc II). The bands for the PCR products are schematically shown. (C) Poly (A)+RNA was extracted from the fat body. Upper lanes: these RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers TGM2F and TGM2R. Lower lanes: these RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers BmA3QPCR1F and BmA3QPCR1R. Resulting products were separated on a 1% agarose gel and visualized with SYBR Green I (Molecular Probes) at a dilution of 1: 10,000 (upper lanes) and visualized with ethidium bromide (lower lanes). M represents the DNA marker (λ /Hind III+ φX174/ Hinc II). The bands for the PCR products are schematically shown.

    Journal: Journal of Insect Science

    Article Title: The Bmdsx transgene including trimmed introns is sex-specifically spliced in tissues of the silkworm, Bombyx mori

    doi:

    Figure Lengend Snippet: Pre-mRNA transcribed from the Bmdsx mini gene is sex-specifically spliced. (A) The diagram shows the structure of the Bmdsx minigene. Open boxes: common exons. Shaded boxes: female-specific exons. Arrows: primers for PCR. (B) Poly (A)+RNA was extracted from the fat body of transgenic silkworms. These RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers TGM2F and TGM2R. Resulting products were separated on a 1% agarose gel and visualized with SYBR Green I (Molecular Probes) at a dilution of 1: 10,000. M represents the DNA marker(λ /Hind III+ φX174/ Hinc II). The bands for the PCR products are schematically shown. (C) Poly (A)+RNA was extracted from the fat body. Upper lanes: these RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers TGM2F and TGM2R. Lower lanes: these RNAs were reverse-transcribed with random hexamer, and the cDNAs were PCR-amplified with primers BmA3QPCR1F and BmA3QPCR1R. Resulting products were separated on a 1% agarose gel and visualized with SYBR Green I (Molecular Probes) at a dilution of 1: 10,000 (upper lanes) and visualized with ethidium bromide (lower lanes). M represents the DNA marker (λ /Hind III+ φX174/ Hinc II). The bands for the PCR products are schematically shown.

    Article Snippet: RT-PCR was performed using the LA RNA PCR kit (Takara, www.takara-bio.co.jp ) following the manufacturer's instructions. cDNA was produced by random priming.

    Techniques: Polymerase Chain Reaction, Transgenic Assay, Random Hexamer Labeling, Amplification, Agarose Gel Electrophoresis, SYBR Green Assay, Marker