la 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
    Buy from Supplier


    Structured Review

    TaKaRa la pcr kit
    Identification of the mutant strain RA1062. A <t>PCR</t> amplification. M: <t>Takara</t> 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
    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/la pcr kit/product/TaKaRa
    Average 99 stars, based on 37927 article reviews
    Price from $9.99 to $1999.99
    la pcr kit - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "The Riemerella anatipestifer M949_RS01035 gene is involved in bacterial lipopolysaccharide biosynthesis"

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

    Journal: Veterinary Research

    doi: 10.1186/s13567-018-0589-8

    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
    Figure Legend 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

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

    2) Product Images from "Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification"

    Article Title: Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.39.9.3326-3331.2001

    Broad-range PCR-RFLP of groESL genes among eight enterococcal species. Lane M, DNA size markers. Lanes 1 to 8, Rsa I digestion; lanes 10 to 16, Hae III. Lanes 1 and 9, E. faecalis ; lanes 2 and 10, E. faecium ; lanes 3 and 11, E. casseliflavus ; lanes 4 and 12, E. gallinarum ; lanes 5 and 13, E. avium ; lanes 6 and 14, E. raffinosus ; lanes 7 and 15, E. durans ; and lanes 8 and 16, E. hirae .
    Figure Legend Snippet: Broad-range PCR-RFLP of groESL genes among eight enterococcal species. Lane M, DNA size markers. Lanes 1 to 8, Rsa I digestion; lanes 10 to 16, Hae III. Lanes 1 and 9, E. faecalis ; lanes 2 and 10, E. faecium ; lanes 3 and 11, E. casseliflavus ; lanes 4 and 12, E. gallinarum ; lanes 5 and 13, E. avium ; lanes 6 and 14, E. raffinosus ; lanes 7 and 15, E. durans ; and lanes 8 and 16, E. hirae .

    Techniques Used: Polymerase Chain Reaction

    3) Product Images from "Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR"

    Article Title: Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0207071

    Different radio-sensitivity of C . neoformans H99S vs H99 strains and their DNA damage progression after 1 kGy exposure. (A) H99S and H99 cells were cultured in YPD overnight, and then 10-fold serially diluted (10 2 to 10 5 ) in PBS and 3 μL of diluted solution was spotted onto the YPD plates. Cells were exposed to the indicated doses of γ-radiation and then further incubated at 30 ° C for 5 days. Image was from one representative experiment of three independent experiments. (B) (C) DNA damage progression of C . neoformans H99S and H99 cells after 1 kGy γ-radiation measured by mitochondrial DNA and nuclear DNA LR-QPCR, respectively. After radiation, cells were harvested at selected time points and DNA was extracted and PCR was performed using 1 ng DNA and 30 ng DNA for the long mitochondrial DNA fragment and long nuclear DNA fragment respectively. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p
    Figure Legend Snippet: Different radio-sensitivity of C . neoformans H99S vs H99 strains and their DNA damage progression after 1 kGy exposure. (A) H99S and H99 cells were cultured in YPD overnight, and then 10-fold serially diluted (10 2 to 10 5 ) in PBS and 3 μL of diluted solution was spotted onto the YPD plates. Cells were exposed to the indicated doses of γ-radiation and then further incubated at 30 ° C for 5 days. Image was from one representative experiment of three independent experiments. (B) (C) DNA damage progression of C . neoformans H99S and H99 cells after 1 kGy γ-radiation measured by mitochondrial DNA and nuclear DNA LR-QPCR, respectively. After radiation, cells were harvested at selected time points and DNA was extracted and PCR was performed using 1 ng DNA and 30 ng DNA for the long mitochondrial DNA fragment and long nuclear DNA fragment respectively. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Techniques Used: Cell Culture, Incubation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p
    Figure Legend Snippet: Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Techniques Used: Irradiation, DNA Extraction, Polymerase Chain Reaction, Picogreen Assay, Amplification

    Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.
    Figure Legend Snippet: Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.

    Techniques Used: Amplification, Real-time Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Picogreen Assay

    4) Product Images from "Loss of Heterozygosity Analysis Using Whole Genome Amplification, Cell Sorting, and Fluorescence-Based PCR"

    Article Title: Loss of Heterozygosity Analysis Using Whole Genome Amplification, Cell Sorting, and Fluorescence-Based PCR

    Journal: Genome Research

    doi:

    PEP preserves relative allele concentrations. Two cases were selected that were informative at the chromosome 17 STR D17S1303, and that had one common allele (diamonds) of the same size (bp), and one unique allele of different size [NL1 (triangles) or NL2 (squares) allele]. Locus-specific reactions were performed with either genomic DNA from the equivalent of 3000 cells (open symbols) or pools of three independent PEP reactions (1000 cells into each PEP) (solid symbols). DNA was mixed with the indicated proportions of NL1:NL2 prior to PEP or locus-specific genomic PCR. The percentage of PCR product represented by each allele was determined by dividing the fluorescent intensity of the allele by the total fluorescent units for all three alleles.
    Figure Legend Snippet: PEP preserves relative allele concentrations. Two cases were selected that were informative at the chromosome 17 STR D17S1303, and that had one common allele (diamonds) of the same size (bp), and one unique allele of different size [NL1 (triangles) or NL2 (squares) allele]. Locus-specific reactions were performed with either genomic DNA from the equivalent of 3000 cells (open symbols) or pools of three independent PEP reactions (1000 cells into each PEP) (solid symbols). DNA was mixed with the indicated proportions of NL1:NL2 prior to PEP or locus-specific genomic PCR. The percentage of PCR product represented by each allele was determined by dividing the fluorescent intensity of the allele by the total fluorescent units for all three alleles.

    Techniques Used: Polymerase Chain Reaction

    LOH detection in the presence of normal cell contamination. Two DNA samples, one corresponding to a biopsy sample with no allelic loss on chromosome 18 (diploid), the other corresponding to a biopsy sample that had undergone chromosome 18 nondisjunction (aneuploid), underwent PEP reactions with genomic input DNA from 1000 cells. Pooled PEP reactions were mixed in the ratios indicated and subjected to locus-specific PCR. ( a ) Electropherograms showing allele ratios in diploid/aneuploid mixtures for four chromosome 18 STRs. Numbers indicate allele ratios. The reciprocal of the allele ratio was taken for those STRs that had ratios > 1 (i.e., larger allele was lost), for ease of comparison. ( b ) Variable STR sensitivity for LOH detection. QLOH for each sample was determined by dividing the allele ratio obtained for each DNA mixture by the allele ratio of the all-diploid sample. The STRs that were most sensitive (shaded squares) and least sensitive (solid diamonds) in detecting a minority population of cells with LOH correspond to the upper (D18S1369) and lower (D18S541) plots, respectively. The average plot (triangles) represents the numerical average of QLOH for all 14 informative markers; the expected plot (line) represents a theoretical line assuming the allele intensity is exactly proportional to its abundance in a population. QLOH was corrected for the contribution of stutter for samples in which the lost allele was in the stutter position.
    Figure Legend Snippet: LOH detection in the presence of normal cell contamination. Two DNA samples, one corresponding to a biopsy sample with no allelic loss on chromosome 18 (diploid), the other corresponding to a biopsy sample that had undergone chromosome 18 nondisjunction (aneuploid), underwent PEP reactions with genomic input DNA from 1000 cells. Pooled PEP reactions were mixed in the ratios indicated and subjected to locus-specific PCR. ( a ) Electropherograms showing allele ratios in diploid/aneuploid mixtures for four chromosome 18 STRs. Numbers indicate allele ratios. The reciprocal of the allele ratio was taken for those STRs that had ratios > 1 (i.e., larger allele was lost), for ease of comparison. ( b ) Variable STR sensitivity for LOH detection. QLOH for each sample was determined by dividing the allele ratio obtained for each DNA mixture by the allele ratio of the all-diploid sample. The STRs that were most sensitive (shaded squares) and least sensitive (solid diamonds) in detecting a minority population of cells with LOH correspond to the upper (D18S1369) and lower (D18S541) plots, respectively. The average plot (triangles) represents the numerical average of QLOH for all 14 informative markers; the expected plot (line) represents a theoretical line assuming the allele intensity is exactly proportional to its abundance in a population. QLOH was corrected for the contribution of stutter for samples in which the lost allele was in the stutter position.

    Techniques Used: Polymerase Chain Reaction

    5) Product Images from "Functional Analysis of Fructosyl-Amino Acid Oxidases of Aspergillus oryzae"

    Article Title: Functional Analysis of Fructosyl-Amino Acid Oxidases of Aspergillus oryzae

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.70.10.5882-5890.2004

    Amplification of partial FAO genes with an FAOD-specific primer set (A) and FAO cDNAs for expression in E. coli (B). In panel A, the FAOD-specific primer set was designed from the alignment of fungal FAODs reported previously. The nucleotide positions of these FAODs for 5′- and 3′-primer designs were as follows: FAOD of A. terreus at positions 784 to 806 and 997 to 1016, amadoriase I of A. fumigatus at positions 797 to 819 and 1016 to 1035, amadoriase II of A. fumigatus at positions 787 to 809 and 1000 to 1019; FOD of F. oxysporum at positions 796 to 818 and 1012 to 1031. Lanes in panel A: 1, φX174-HincII digest; 2, PCR product. Lanes in panel B: 1, λ-EcoT14 I digest; 2, FAOAo1 ; 3, FAOAo2 .
    Figure Legend Snippet: Amplification of partial FAO genes with an FAOD-specific primer set (A) and FAO cDNAs for expression in E. coli (B). In panel A, the FAOD-specific primer set was designed from the alignment of fungal FAODs reported previously. The nucleotide positions of these FAODs for 5′- and 3′-primer designs were as follows: FAOD of A. terreus at positions 784 to 806 and 997 to 1016, amadoriase I of A. fumigatus at positions 797 to 819 and 1016 to 1035, amadoriase II of A. fumigatus at positions 787 to 809 and 1000 to 1019; FOD of F. oxysporum at positions 796 to 818 and 1012 to 1031. Lanes in panel A: 1, φX174-HincII digest; 2, PCR product. Lanes in panel B: 1, λ-EcoT14 I digest; 2, FAOAo1 ; 3, FAOAo2 .

    Techniques Used: Amplification, Expressing, Polymerase Chain Reaction

    6) Product Images from "MzPIP2;1: An Aquaporin Involved in Radial Water Movement in Both Water Uptake and Transportation, Altered the Drought and Salt Tolerance of Transgenic Arabidopsis"

    Article Title: MzPIP2;1: An Aquaporin Involved in Radial Water Movement in Both Water Uptake and Transportation, Altered the Drought and Salt Tolerance of Transgenic Arabidopsis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0142446

    The phenotype of young transgenic Arabidopsis plants ectopically expressing MzPIP2;1 under salt stress. (A) The expression levels of MzPIP2;1 in three transgenic lines and wild-type by semi-quantitative RT-PCR, with the AtActin as internal gene; (B)The phenotype of three transgenic lines and wild-type under 100 NaCl and 130mM NaCl treatments. WT: wild-type.
    Figure Legend Snippet: The phenotype of young transgenic Arabidopsis plants ectopically expressing MzPIP2;1 under salt stress. (A) The expression levels of MzPIP2;1 in three transgenic lines and wild-type by semi-quantitative RT-PCR, with the AtActin as internal gene; (B)The phenotype of three transgenic lines and wild-type under 100 NaCl and 130mM NaCl treatments. WT: wild-type.

    Techniques Used: Transgenic Assay, Expressing, Quantitative RT-PCR

    7) Product Images from "Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples"

    Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-30322-y

    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .
    Figure Legend Snippet: ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Produced, Diagnostic Assay

    8) Product Images from "Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR"

    Article Title: Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0207071

    Different radio-sensitivity of C . neoformans H99S vs H99 strains and their DNA damage progression after 1 kGy exposure. (A) H99S and H99 cells were cultured in YPD overnight, and then 10-fold serially diluted (10 2 to 10 5 ) in PBS and 3 μL of diluted solution was spotted onto the YPD plates. Cells were exposed to the indicated doses of γ-radiation and then further incubated at 30 ° C for 5 days. Image was from one representative experiment of three independent experiments. (B) (C) DNA damage progression of C . neoformans H99S and H99 cells after 1 kGy γ-radiation measured by mitochondrial DNA and nuclear DNA LR-QPCR, respectively. After radiation, cells were harvested at selected time points and DNA was extracted and PCR was performed using 1 ng DNA and 30 ng DNA for the long mitochondrial DNA fragment and long nuclear DNA fragment respectively. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p
    Figure Legend Snippet: Different radio-sensitivity of C . neoformans H99S vs H99 strains and their DNA damage progression after 1 kGy exposure. (A) H99S and H99 cells were cultured in YPD overnight, and then 10-fold serially diluted (10 2 to 10 5 ) in PBS and 3 μL of diluted solution was spotted onto the YPD plates. Cells were exposed to the indicated doses of γ-radiation and then further incubated at 30 ° C for 5 days. Image was from one representative experiment of three independent experiments. (B) (C) DNA damage progression of C . neoformans H99S and H99 cells after 1 kGy γ-radiation measured by mitochondrial DNA and nuclear DNA LR-QPCR, respectively. After radiation, cells were harvested at selected time points and DNA was extracted and PCR was performed using 1 ng DNA and 30 ng DNA for the long mitochondrial DNA fragment and long nuclear DNA fragment respectively. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Techniques Used: Cell Culture, Incubation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p
    Figure Legend Snippet: Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Techniques Used: Irradiation, DNA Extraction, Polymerase Chain Reaction, Picogreen Assay, Amplification

    Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.
    Figure Legend Snippet: Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.

    Techniques Used: Amplification, Real-time Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Picogreen Assay

    9) Product Images from "Development of a novel detection system for microbes from bovine diarrhea by real-time PCR"

    Article Title: Development of a novel detection system for microbes from bovine diarrhea by real-time PCR

    Journal: The Journal of Veterinary Medical Science

    doi: 10.1292/jvms.15-0552

    Dembo-PCR workflow. To prepare each sample for assay, 10% fecal suspensions were made in PBS (−). The suspensions were then used directly for the extraction of bacteria and protozoa nucleic acids with a QIAamp Fast DNA Stool Mini Kit. For virus detection, the suspensions were centrifuged for 15 min at 10,000 rpm, and viral DNA and RNA were extracted from the supernatants with a High Pure Viral Nucleic Acid Kit. After pathogen RNA and DNA were extracted, samples, reagents and each primer and probe were mixed in individual reaction tubes. Samples were applied at 2 µl per tube. A LightCycler Nano was used for all qPCR reactions performed in this study. A one step PrimeScript RT-PCR Kit (Perfect Real time) was used for amplification of extracts from RNA viruses, and Premix Ex Taq (Perfect Real time) was used for amplification of extracts from DNA viruses, bacteria and protozoa.
    Figure Legend Snippet: Dembo-PCR workflow. To prepare each sample for assay, 10% fecal suspensions were made in PBS (−). The suspensions were then used directly for the extraction of bacteria and protozoa nucleic acids with a QIAamp Fast DNA Stool Mini Kit. For virus detection, the suspensions were centrifuged for 15 min at 10,000 rpm, and viral DNA and RNA were extracted from the supernatants with a High Pure Viral Nucleic Acid Kit. After pathogen RNA and DNA were extracted, samples, reagents and each primer and probe were mixed in individual reaction tubes. Samples were applied at 2 µl per tube. A LightCycler Nano was used for all qPCR reactions performed in this study. A one step PrimeScript RT-PCR Kit (Perfect Real time) was used for amplification of extracts from RNA viruses, and Premix Ex Taq (Perfect Real time) was used for amplification of extracts from DNA viruses, bacteria and protozoa.

    Techniques Used: Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Amplification

    10) Product Images from "Influence of simulated microgravity on the activation of the small GTPase Rho involved in cytoskeletal formation - molecular cloning and sequencing of bovine leukemia-associated guanine nucleotide exchange factor"

    Article Title: Influence of simulated microgravity on the activation of the small GTPase Rho involved in cytoskeletal formation - molecular cloning and sequencing of bovine leukemia-associated guanine nucleotide exchange factor

    Journal: BMC Biochemistry

    doi: 10.1186/1471-2091-7-19

    Real-time quantitative PCR . The expression of LARG mRNA was measured by real-time quantitative PCR as described in the Methods section. The LARG gene expression in the clinorotated cells decreased to 57.5% of that in the stationary control cells. Values were obtained from seven individual experiments in triplicates and are represented as means ± SE ( p
    Figure Legend Snippet: Real-time quantitative PCR . The expression of LARG mRNA was measured by real-time quantitative PCR as described in the Methods section. The LARG gene expression in the clinorotated cells decreased to 57.5% of that in the stationary control cells. Values were obtained from seven individual experiments in triplicates and are represented as means ± SE ( p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    11) Product Images from "Novel Plasmid-Mediated 16S rRNA Methylase, RmtC, Found in a Proteus mirabilis Isolate Demonstrating Extraordinary High-Level Resistance against Various Aminoglycosides"

    Article Title: Novel Plasmid-Mediated 16S rRNA Methylase, RmtC, Found in a Proteus mirabilis Isolate Demonstrating Extraordinary High-Level Resistance against Various Aminoglycosides

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.50.1.178-184.2006

    (A) Schematic presentation of the 7.7-kb EcoRI fragment on pBC-E1 and the 1.2-kb PCR fragment on pBC-KB1. (B) Part of the nucleotide sequences encoding the 3′ end of an IS Ecp1 -like element and the start region of rmtC . The predicted −35 and −10 promoter sequences and the +1 position of the putative transcriptional start of rmtC ). Arrows indicate the transcription orientation. The deduced amino acid sequences are designated in single-letter code. The right inverted repeat (IR) of an IS Ecp1 -like element is underlined.
    Figure Legend Snippet: (A) Schematic presentation of the 7.7-kb EcoRI fragment on pBC-E1 and the 1.2-kb PCR fragment on pBC-KB1. (B) Part of the nucleotide sequences encoding the 3′ end of an IS Ecp1 -like element and the start region of rmtC . The predicted −35 and −10 promoter sequences and the +1 position of the putative transcriptional start of rmtC ). Arrows indicate the transcription orientation. The deduced amino acid sequences are designated in single-letter code. The right inverted repeat (IR) of an IS Ecp1 -like element is underlined.

    Techniques Used: Polymerase Chain Reaction

    12) Product Images from "Lack of Parkin Anticipates the Phenotype and Affects Mitochondrial Morphology and mtDNA Levels in a Mouse Model of Parkinson's Disease"

    Article Title: Lack of Parkin Anticipates the Phenotype and Affects Mitochondrial Morphology and mtDNA Levels in a Mouse Model of Parkinson's Disease

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1384-17.2017

    mtDNA deletions and mutations increase when endogenous Parkin is lost in striata of PD-mito- Pst I mice. A , Schematic representation of mouse mtDNA. Scissors indicate the Pst I cleavage sites. Red arrows represent encoded proteins. Gray arrows represent mt-tRNAs. Color-coded representation of the position of the amplicons used for the qPCRs is maintained in the graphs. B , mtDNA levels measured by qPCR of LCM-collected SN TH + neurons ( n = 5/group; T = p value after t test). C , D , Southern blot probing striatal DNA with mtDNA and nuclear DNA probes ( C ) and mtDNA content measured by qPCR ( D ) in 4-month-old animals. E , Quantification by qPCR of full-length mtDNA content in striata of 4-month-old animals. F , Recombination events detected by qPCR using primers flanking the Pst I sites in striatal samples. G , H , Long-range PCR amplification of a 10 kb and a 117 bp fragment from striatal DNA ( G ) and relative quantification of a 10 kb fragment normalized to the 117 bp fragment ( H ). Boxes in graphs B , D , and F represent one individual animal. * p
    Figure Legend Snippet: mtDNA deletions and mutations increase when endogenous Parkin is lost in striata of PD-mito- Pst I mice. A , Schematic representation of mouse mtDNA. Scissors indicate the Pst I cleavage sites. Red arrows represent encoded proteins. Gray arrows represent mt-tRNAs. Color-coded representation of the position of the amplicons used for the qPCRs is maintained in the graphs. B , mtDNA levels measured by qPCR of LCM-collected SN TH + neurons ( n = 5/group; T = p value after t test). C , D , Southern blot probing striatal DNA with mtDNA and nuclear DNA probes ( C ) and mtDNA content measured by qPCR ( D ) in 4-month-old animals. E , Quantification by qPCR of full-length mtDNA content in striata of 4-month-old animals. F , Recombination events detected by qPCR using primers flanking the Pst I sites in striatal samples. G , H , Long-range PCR amplification of a 10 kb and a 117 bp fragment from striatal DNA ( G ) and relative quantification of a 10 kb fragment normalized to the 117 bp fragment ( H ). Boxes in graphs B , D , and F represent one individual animal. * p

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Laser Capture Microdissection, Southern Blot, Polymerase Chain Reaction, Amplification

    13) Product Images from "Clg2p interacts with Clf and ClUrase to regulate appressorium formation, pathogenicity and conidial morphology in Curvularia lunata"

    Article Title: Clg2p interacts with Clf and ClUrase to regulate appressorium formation, pathogenicity and conidial morphology in Curvularia lunata

    Journal: Scientific Reports

    doi: 10.1038/srep24047

    Construction and confirmation of the Clg2p deletion mutant and complementation strain. ( A ) Construction of the deletion vector lacking Clg2p . ( B ) Construction of the complementation vector of Clg2p as described in the methods section. ( C,D ) Southern blotting analysis of the Clg2p deletion mutant (Δ Clg2p ) and complementation strain ( Clg2p- Com). ( C ) Southern blotting was performed on isolated total genomic DNA from CX-3, Δ Clg2p and Clg2p- Com after digestion with Xba I and Bam HI. A 607-bp PCR fragment of hph was used as the hybridization probe. ( D ) A second Southern blotting analysis was performed using isolated total genomic DNA from wild-type strain CX-3, Δ Clg2p and Clg2p- Com after digestion with a single-cutter restriction enzyme, Hin dIII, using a 951-bp PCR fragment of Clg2p as the probe. ( E ) Clg2p expression in CX-3, Δ Clg2p and Clg2p- Com was analysed by RT-PCR using the Clg2p gene-specific primers clg2pds and clg2pda. A predicted 415-bp fragment was obtained from the wild-type CX-3 strain and the complemented strain but was absent in the Δ Clg2p mutant.
    Figure Legend Snippet: Construction and confirmation of the Clg2p deletion mutant and complementation strain. ( A ) Construction of the deletion vector lacking Clg2p . ( B ) Construction of the complementation vector of Clg2p as described in the methods section. ( C,D ) Southern blotting analysis of the Clg2p deletion mutant (Δ Clg2p ) and complementation strain ( Clg2p- Com). ( C ) Southern blotting was performed on isolated total genomic DNA from CX-3, Δ Clg2p and Clg2p- Com after digestion with Xba I and Bam HI. A 607-bp PCR fragment of hph was used as the hybridization probe. ( D ) A second Southern blotting analysis was performed using isolated total genomic DNA from wild-type strain CX-3, Δ Clg2p and Clg2p- Com after digestion with a single-cutter restriction enzyme, Hin dIII, using a 951-bp PCR fragment of Clg2p as the probe. ( E ) Clg2p expression in CX-3, Δ Clg2p and Clg2p- Com was analysed by RT-PCR using the Clg2p gene-specific primers clg2pds and clg2pda. A predicted 415-bp fragment was obtained from the wild-type CX-3 strain and the complemented strain but was absent in the Δ Clg2p mutant.

    Techniques Used: Mutagenesis, Plasmid Preparation, Southern Blot, Isolation, Polymerase Chain Reaction, Hybridization, Expressing, Reverse Transcription Polymerase Chain Reaction

    Molecular characteristics of Clg2p in C. lunata. ( A ) Analysis of conserved domains of Clg2p . Box1, box3, box4 and box5 are the GTP/GDP domains; box2 is a binding site for downstream effector molecules (RA); and box6 is a variable region containing a CAAX motif. ( B ) A phylogenetic tree representing the phylogenetic relationships of Clg2p proteins and Ras proteins among related fungi. The number for each interior branch was the percentage of the bootstrap value (1000 replicates). ( C ) Analysis of copy number of Clg2p in the genome. A 951-bp PCR fragment amplified with primers 1F/R as a template of strain CX-3 DNA was labelled using Biotin to make the probe. ( D ) Expression patterns of Clg2p by qRT-PCR. M and C (x-axis) represent mycelial growth in potato dextrose (PD) medium for 3 d and conidia collected from 7 d culture on PDA plates at 28 °C in the dark, respectively. On the graph, 3 h, 6 h and 9 h (x-axis) represent germinating conidia collected from cellophane overlaid on PDA plates at 25 °C after 3, 6 and 9 h of growth, respectively. The error bars were calculated based on three replicates.
    Figure Legend Snippet: Molecular characteristics of Clg2p in C. lunata. ( A ) Analysis of conserved domains of Clg2p . Box1, box3, box4 and box5 are the GTP/GDP domains; box2 is a binding site for downstream effector molecules (RA); and box6 is a variable region containing a CAAX motif. ( B ) A phylogenetic tree representing the phylogenetic relationships of Clg2p proteins and Ras proteins among related fungi. The number for each interior branch was the percentage of the bootstrap value (1000 replicates). ( C ) Analysis of copy number of Clg2p in the genome. A 951-bp PCR fragment amplified with primers 1F/R as a template of strain CX-3 DNA was labelled using Biotin to make the probe. ( D ) Expression patterns of Clg2p by qRT-PCR. M and C (x-axis) represent mycelial growth in potato dextrose (PD) medium for 3 d and conidia collected from 7 d culture on PDA plates at 28 °C in the dark, respectively. On the graph, 3 h, 6 h and 9 h (x-axis) represent germinating conidia collected from cellophane overlaid on PDA plates at 25 °C after 3, 6 and 9 h of growth, respectively. The error bars were calculated based on three replicates.

    Techniques Used: Binding Assay, Polymerase Chain Reaction, Amplification, Expressing, Quantitative RT-PCR

    14) Product Images from "Ferric Enterochelin Transport in Yersinia enterocolitica: Molecular and Evolutionary Aspects"

    Article Title: Ferric Enterochelin Transport in Yersinia enterocolitica: Molecular and Evolutionary Aspects

    Journal: Journal of Bacteriology

    doi:

    Southern blotting of Cla I-digested chromosomal DNA of different Y. enterocolitica serovars and E. coli DH5α. The preparation was probed with a labelled PCR product generated from the Yersinia fes gene. Lane 1, E. coli DH5α; lane 2, Y. enterocolitica O3; lane 3, O5; lane 4, O5.27; lane 5, O8; lane 6, O9; lane 7, O13; lane 8, O20; lane 9, Y. pseudotuberculosis I; lane 10, Y. pseudotuberculosis II; lane 11, Y. pseudotuberculosis III; lane 12, Y. pestis pgm + .
    Figure Legend Snippet: Southern blotting of Cla I-digested chromosomal DNA of different Y. enterocolitica serovars and E. coli DH5α. The preparation was probed with a labelled PCR product generated from the Yersinia fes gene. Lane 1, E. coli DH5α; lane 2, Y. enterocolitica O3; lane 3, O5; lane 4, O5.27; lane 5, O8; lane 6, O9; lane 7, O13; lane 8, O20; lane 9, Y. pseudotuberculosis I; lane 10, Y. pseudotuberculosis II; lane 11, Y. pseudotuberculosis III; lane 12, Y. pestis pgm + .

    Techniques Used: Southern Blot, Polymerase Chain Reaction, Generated

    15) Product Images from "An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody"

    Article Title: An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody

    Journal: BMC Biotechnology

    doi: 10.1186/1472-6750-9-99

    DNA fingerprint analysis of specific scFv genes of positive clones . scFv inserts were amplified from individual colonies of all positive clones. Amplification was done with Ex-Taq DNA polymearase enzyme using pCANTAB5-R1 and pCANTAB5-S6 primers. The amplified products were then analyzed on agarose gels. A . scFv encoding inserts of DNA of 40 diversified positive clones B . Only five restriction patterns of scFv encoding inserts of DNA were identified against five different clones (scFv K1 - K5). Molecular weights of DNA ladder markers (kbp) were indicated by M on the left.
    Figure Legend Snippet: DNA fingerprint analysis of specific scFv genes of positive clones . scFv inserts were amplified from individual colonies of all positive clones. Amplification was done with Ex-Taq DNA polymearase enzyme using pCANTAB5-R1 and pCANTAB5-S6 primers. The amplified products were then analyzed on agarose gels. A . scFv encoding inserts of DNA of 40 diversified positive clones B . Only five restriction patterns of scFv encoding inserts of DNA were identified against five different clones (scFv K1 - K5). Molecular weights of DNA ladder markers (kbp) were indicated by M on the left.

    Techniques Used: Clone Assay, Amplification

    16) Product Images from "The Riemerella anatipestifer M949_RS01035 gene is involved in bacterial lipopolysaccharide biosynthesis"

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

    Journal: Veterinary Research

    doi: 10.1186/s13567-018-0589-8

    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
    Figure Legend 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

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

    17) Product Images from "Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples"

    Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-30322-y

    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .
    Figure Legend Snippet: ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .

    Techniques Used: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Produced, Diagnostic Assay

    18) Product Images from "Sequence Analysis, Overexpression, and Antisense Inhibition of a ?-Xylosidase Gene, xylA, from Aspergillus oryzae KBN616"

    Article Title: Sequence Analysis, Overexpression, and Antisense Inhibition of a ?-Xylosidase Gene, xylA, from Aspergillus oryzae KBN616

    Journal: Applied and Environmental Microbiology

    doi:

    Restriction map of the xylA gene. The sequencing strategy for the xylA gene is represented below the restriction map by arrows. Solid and hatched boxes indicate the coding regions of the xylA gene and the BXF1 fragment amplified by PCR, respectively.
    Figure Legend Snippet: Restriction map of the xylA gene. The sequencing strategy for the xylA gene is represented below the restriction map by arrows. Solid and hatched boxes indicate the coding regions of the xylA gene and the BXF1 fragment amplified by PCR, respectively.

    Techniques Used: Sequencing, Amplification, Polymerase Chain Reaction

    19) Product Images from "Cloning and Molecular Analyses of a Gibberellin 20-Oxidase Gene Expressed Specifically in Developing Seeds of Watermelon 1"

    Article Title: Cloning and Molecular Analyses of a Gibberellin 20-Oxidase Gene Expressed Specifically in Developing Seeds of Watermelon 1

    Journal: Plant Physiology

    doi:

    Genomic DNA-blot analysis of Cv20ox . Ten micrograms of watermelon genomic DNA was digested with Eco RI (E) or Hin dIII (H) and resolved on a 0.8% agarose gel. The 300-bp cDNA cloned by PCR was used as a probe. The positions of Hin dIII-digested λ-DNA size markers are shown on the right in kb.
    Figure Legend Snippet: Genomic DNA-blot analysis of Cv20ox . Ten micrograms of watermelon genomic DNA was digested with Eco RI (E) or Hin dIII (H) and resolved on a 0.8% agarose gel. The 300-bp cDNA cloned by PCR was used as a probe. The positions of Hin dIII-digested λ-DNA size markers are shown on the right in kb.

    Techniques Used: Agarose Gel Electrophoresis, Clone Assay, Polymerase Chain Reaction

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    Synthesized:

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    Reverse Transcription Polymerase Chain Reaction:

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    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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    TaKaRa la taq dna polymerase
    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying <t>DNA</t> polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA <t>Taq</t> (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .
    La Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 656 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/la taq dna polymerase/product/TaKaRa
    Average 99 stars, based on 656 article reviews
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    89
    TaKaRa la taq enzyme
    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying <t>DNA</t> polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA <t>Taq</t> (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .
    La Taq Enzyme, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/la taq enzyme/product/TaKaRa
    Average 89 stars, based on 8 article reviews
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    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .

    Journal: Scientific Reports

    Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples

    doi: 10.1038/s41598-018-30322-y

    Figure Lengend Snippet: ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 O control. Full-length gels are presented in Supplementary Fig. 1 .

    Article Snippet: The following enzymes were tested: Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech).

    Techniques: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Produced, Diagnostic Assay

    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .

    Journal: Scientific Reports

    Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples

    doi: 10.1038/s41598-018-30322-y

    Figure Lengend Snippet: ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .

    Article Snippet: The following enzymes were tested: Takara LA Taq DNA Polymerase (Clontech), AccuTaq LA DNA Polymerase (Sigma) and PrimeSTAR GXL DNA Polymerase (Clontech).

    Techniques: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Produced, Diagnostic Assay