multiple pcrs  (Qiagen)

 
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    Name:
    HotStarTaq Master Mix Kit
    Description:
    For highly specific amplification for any PCR application Kit contents Qiagen HotStarTaq Master Mix Kit 250U 5U L 10 min at 97C 60 min at 94C Half life Genomic DNA and cDNA Sample Recombinant Enzyme PCR Amplification Ready to use Ideal for Highly Specific Amplification for any PCR Application Includes 3 x 0 85mL HotStarTaq Master Mix Contains 250U HotStarTaq DNA Polymerase PCR Buffer with 3mM MgCl2 and 400mM of Each dNTP and 2 x 1 7mL RNase free Water Benefits High PCR specificity without the need for optimization Easy reaction setup at room temperature Ready to use master mix format reduces pipetting step
    Catalog Number:
    203443
    Price:
    220
    Category:
    HotStarTaq Master Mix Kit
    Buy from Supplier


    Structured Review

    Qiagen multiple pcrs
    HotStarTaq Master Mix Kit
    For highly specific amplification for any PCR application Kit contents Qiagen HotStarTaq Master Mix Kit 250U 5U L 10 min at 97C 60 min at 94C Half life Genomic DNA and cDNA Sample Recombinant Enzyme PCR Amplification Ready to use Ideal for Highly Specific Amplification for any PCR Application Includes 3 x 0 85mL HotStarTaq Master Mix Contains 250U HotStarTaq DNA Polymerase PCR Buffer with 3mM MgCl2 and 400mM of Each dNTP and 2 x 1 7mL RNase free Water Benefits High PCR specificity without the need for optimization Easy reaction setup at room temperature Ready to use master mix format reduces pipetting step
    https://www.bioz.com/result/multiple pcrs/product/Qiagen
    Average 85 stars, based on 36704 article reviews
    Price from $9.99 to $1999.99
    multiple pcrs - by Bioz Stars, 2020-08
    85/100 stars

    Images

    1) Product Images from "Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis"

    Article Title: Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis

    Journal: Experimental Animals

    doi: 10.1538/expanim.62.267

    Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.
    Figure Legend Snippet: Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.

    Techniques Used: Mouse Assay, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

    2) Product Images from "Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis"

    Article Title: Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis

    Journal: Experimental Animals

    doi: 10.1538/expanim.62.267

    Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.
    Figure Legend Snippet: Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.

    Techniques Used: Mouse Assay, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

    3) Product Images from "Stress exposure and psychopathology alter methylation of the serotonin receptor 2A (HTR2A) gene in preschoolers"

    Article Title: Stress exposure and psychopathology alter methylation of the serotonin receptor 2A (HTR2A) gene in preschoolers

    Journal: Development and psychopathology

    doi: 10.1017/S0954579417001274

    Mean differences in methylation by HTR2A genotype Note: Different letters indicate significant differences within the CpG site based on genotype at p
    Figure Legend Snippet: Mean differences in methylation by HTR2A genotype Note: Different letters indicate significant differences within the CpG site based on genotype at p

    Techniques Used: Methylation

    HTR2A genotype moderates the association of contextual stress and HTR2A −1420 methylation
    Figure Legend Snippet: HTR2A genotype moderates the association of contextual stress and HTR2A −1420 methylation

    Techniques Used: Methylation

    4) Product Images from "A human somatic cell culture system for modelling gene silencing by transcriptional interference"

    Article Title: A human somatic cell culture system for modelling gene silencing by transcriptional interference

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2020.e03261

    Expression and methylation analyses in Flp-In T-REx 293. A) Targeting constructs. Dark grey: upstream CMV promoter, light grey: minigene consisting of test promoter (MSH2 , HBA2, UBE3A ) and EGFP , white: exons 2 and 3 of ocHBB2 , triangle: poly(A) signal, black bar: linker for integration of minigenes, arrows: primers for qRT-PCR. B) Expression of ocHBB2 (top) and EGFP (bottom) upon induction with doxycycline, black: 3 days, white 14 days. Short line indicates mean fold induction/reduction, number below indicates fold change relative to uninduced state, dotted line: uninduced set to 1. Symbols: independent clones. C) DNA methylation of test promoters before and after induction (-/+ dox). Blue: not methylated, red: methylated, CGs in columns, reads in rows. Overall percentage of DNA methylation and number of reads are indicated. In comparison to the example plot in Figure 1 B, there are no methylated (red) reads. D) Expression of ocHBB2 (top) and EGFP (bottom) after 3 days induction (dark grey bars) and after 3 days without induction (light grey bars). Numbers below indicate average fold change between these two states. Dotted line: uninduced state set to 1.
    Figure Legend Snippet: Expression and methylation analyses in Flp-In T-REx 293. A) Targeting constructs. Dark grey: upstream CMV promoter, light grey: minigene consisting of test promoter (MSH2 , HBA2, UBE3A ) and EGFP , white: exons 2 and 3 of ocHBB2 , triangle: poly(A) signal, black bar: linker for integration of minigenes, arrows: primers for qRT-PCR. B) Expression of ocHBB2 (top) and EGFP (bottom) upon induction with doxycycline, black: 3 days, white 14 days. Short line indicates mean fold induction/reduction, number below indicates fold change relative to uninduced state, dotted line: uninduced set to 1. Symbols: independent clones. C) DNA methylation of test promoters before and after induction (-/+ dox). Blue: not methylated, red: methylated, CGs in columns, reads in rows. Overall percentage of DNA methylation and number of reads are indicated. In comparison to the example plot in Figure 1 B, there are no methylated (red) reads. D) Expression of ocHBB2 (top) and EGFP (bottom) after 3 days induction (dark grey bars) and after 3 days without induction (light grey bars). Numbers below indicate average fold change between these two states. Dotted line: uninduced state set to 1.

    Techniques Used: Expressing, Methylation, Construct, Quantitative RT-PCR, DNA Methylation Assay

    5) Product Images from "A human somatic cell culture system for modelling gene silencing by transcriptional interference"

    Article Title: A human somatic cell culture system for modelling gene silencing by transcriptional interference

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2020.e03261

    Expression and methylation analyses in Flp-In T-REx 293. A) Targeting constructs. Dark grey: upstream CMV promoter, light grey: minigene consisting of test promoter (MSH2 , HBA2, UBE3A ) and EGFP , white: exons 2 and 3 of ocHBB2 , triangle: poly(A) signal, black bar: linker for integration of minigenes, arrows: primers for qRT-PCR. B) Expression of ocHBB2 (top) and EGFP (bottom) upon induction with doxycycline, black: 3 days, white 14 days. Short line indicates mean fold induction/reduction, number below indicates fold change relative to uninduced state, dotted line: uninduced set to 1. Symbols: independent clones. C) DNA methylation of test promoters before and after induction (-/+ dox). Blue: not methylated, red: methylated, CGs in columns, reads in rows. Overall percentage of DNA methylation and number of reads are indicated. In comparison to the example plot in Figure 1 B, there are no methylated (red) reads. D) Expression of ocHBB2 (top) and EGFP (bottom) after 3 days induction (dark grey bars) and after 3 days without induction (light grey bars). Numbers below indicate average fold change between these two states. Dotted line: uninduced state set to 1.
    Figure Legend Snippet: Expression and methylation analyses in Flp-In T-REx 293. A) Targeting constructs. Dark grey: upstream CMV promoter, light grey: minigene consisting of test promoter (MSH2 , HBA2, UBE3A ) and EGFP , white: exons 2 and 3 of ocHBB2 , triangle: poly(A) signal, black bar: linker for integration of minigenes, arrows: primers for qRT-PCR. B) Expression of ocHBB2 (top) and EGFP (bottom) upon induction with doxycycline, black: 3 days, white 14 days. Short line indicates mean fold induction/reduction, number below indicates fold change relative to uninduced state, dotted line: uninduced set to 1. Symbols: independent clones. C) DNA methylation of test promoters before and after induction (-/+ dox). Blue: not methylated, red: methylated, CGs in columns, reads in rows. Overall percentage of DNA methylation and number of reads are indicated. In comparison to the example plot in Figure 1 B, there are no methylated (red) reads. D) Expression of ocHBB2 (top) and EGFP (bottom) after 3 days induction (dark grey bars) and after 3 days without induction (light grey bars). Numbers below indicate average fold change between these two states. Dotted line: uninduced state set to 1.

    Techniques Used: Expressing, Methylation, Construct, Quantitative RT-PCR, DNA Methylation Assay

    6) Product Images from "Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis"

    Article Title: Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis

    Journal: Experimental Animals

    doi: 10.1538/expanim.62.267

    Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.
    Figure Legend Snippet: Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.

    Techniques Used: Mouse Assay, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

    PCR for genotyping Hr alleles in HR and Hos:HR-1 mice. (A) Primer positions (primer sequences are shown in Table 1 ). PCR using the primers mHR-int6-S607 (S607) and mHR-int6-R850 (R850) produces 244-bp amplicons from wild-type alleles ( Hr , + in Fig. 3B ) only. PCR using the primers mHR-int6-S776 (S776) and mHR-int6-R850 (R850) produces 275-bp amplicons from mutant alleles ( Hr hr , hr in Fig. 3B ) only. (B) Zygosity determination by PCR. Zygosities of Hr alleles were determined by PCR using three primers (S607, S776, and R850) simultaneously. If only 275-bp amplicons were produced, the mice were taken to be homozygous ( Hr hr / Hr hr ). If only 244-bp amplicons were produced, the mice were wild-type ( Hr / Hr ). If both amplicons were produced, the mice were heterozygous ( Hr / Hr hr ). Electropherograms of PCR products indicate the zygosities of homozygous, heterozygous, and wild-type HR mice as well as homozygous Hos:HR-1 mice.
    Figure Legend Snippet: PCR for genotyping Hr alleles in HR and Hos:HR-1 mice. (A) Primer positions (primer sequences are shown in Table 1 ). PCR using the primers mHR-int6-S607 (S607) and mHR-int6-R850 (R850) produces 244-bp amplicons from wild-type alleles ( Hr , + in Fig. 3B ) only. PCR using the primers mHR-int6-S776 (S776) and mHR-int6-R850 (R850) produces 275-bp amplicons from mutant alleles ( Hr hr , hr in Fig. 3B ) only. (B) Zygosity determination by PCR. Zygosities of Hr alleles were determined by PCR using three primers (S607, S776, and R850) simultaneously. If only 275-bp amplicons were produced, the mice were taken to be homozygous ( Hr hr / Hr hr ). If only 244-bp amplicons were produced, the mice were wild-type ( Hr / Hr ). If both amplicons were produced, the mice were heterozygous ( Hr / Hr hr ). Electropherograms of PCR products indicate the zygosities of homozygous, heterozygous, and wild-type HR mice as well as homozygous Hos:HR-1 mice.

    Techniques Used: Polymerase Chain Reaction, Mouse Assay, Mutagenesis, Produced

    7) Product Images from "Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition"

    Article Title: Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-12-178

    a. Detection of spiked KPL-4 cells and cells after i.v. injection. 10 cells were spiked into 100 μl or 1 ml of native murine blood samples. Cells of the KPL-4 line stably expressed EpCAM, Her2 and the housekeeping-genes for up to 4 h. The cells could be detected by AdnaTest and without antibody enrichment (a.). 100,000 cells of the KPL-4 cell line were injected into the tail vein of native mice. Tumour cells could not be detected by the AdnaTest, whereas human mRNA was visible without antibody enrichment for up to 4 h after injection. The detected cells showed serial changes and reduced EpCAM expression already 30 min after i.v. injection (b.). b. Tissue analyses for human mRNA after injection of 100,000 KPL-4 cells into the tail vein. Tissues were homogenised and RNA was purified followed by cDNA synthesis and PCR reaction. Human breast cancer cells detectable in the lungs and one liver were positive for EpCAM and Her2 expression up to 2 h after KPL-4 cell injection. c. MDA-MB-231 breast cancer cells were injected into the heart and blood was analysed for human mRNA signals. CTCs were detectable in all animals for up to 5 h after injection. The detected cells showed changes of mRNA expression including downregulation of EpCAM and MUC-1, whereas mRNA of the housekeeping-genes and mesenchymal Vimentin was visible permanently.
    Figure Legend Snippet: a. Detection of spiked KPL-4 cells and cells after i.v. injection. 10 cells were spiked into 100 μl or 1 ml of native murine blood samples. Cells of the KPL-4 line stably expressed EpCAM, Her2 and the housekeeping-genes for up to 4 h. The cells could be detected by AdnaTest and without antibody enrichment (a.). 100,000 cells of the KPL-4 cell line were injected into the tail vein of native mice. Tumour cells could not be detected by the AdnaTest, whereas human mRNA was visible without antibody enrichment for up to 4 h after injection. The detected cells showed serial changes and reduced EpCAM expression already 30 min after i.v. injection (b.). b. Tissue analyses for human mRNA after injection of 100,000 KPL-4 cells into the tail vein. Tissues were homogenised and RNA was purified followed by cDNA synthesis and PCR reaction. Human breast cancer cells detectable in the lungs and one liver were positive for EpCAM and Her2 expression up to 2 h after KPL-4 cell injection. c. MDA-MB-231 breast cancer cells were injected into the heart and blood was analysed for human mRNA signals. CTCs were detectable in all animals for up to 5 h after injection. The detected cells showed changes of mRNA expression including downregulation of EpCAM and MUC-1, whereas mRNA of the housekeeping-genes and mesenchymal Vimentin was visible permanently.

    Techniques Used: Injection, Stable Transfection, Mouse Assay, Expressing, Purification, Polymerase Chain Reaction, Multiple Displacement Amplification

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    Isolation:

    Article Title: Paradoxical effects of repeat interruptions on spinocerebellar ataxia type 10 expansions and repeat instability
    Article Snippet: .. Genomic DNA isolated from SCA10 somatic cell hybrid lines was used as template in a long-range PCR using HotStarTaq Master Mix (Qiagen, Germantown, MD, USA) with forward (5′- GTCATCAGGTTGTTGCTGTGGAAAGGGGCGGTA-3′) and reverse (5′- GCCACTGCACTCCAGCCTGGGCAACATAGAGAG-3′) primers flanking the repeat. .. PCR products were cloned into the TOPO-TA vector (Invitrogen, Grand Island, NY, USA) and propagated in Escherichia coli grown at 25 °C.

    Cell Culture:

    Article Title: High Frequency of a Novel Filamentous Phage, VCY?, within an Environmental Vibrio cholerae Population
    Article Snippet: .. In a typical reaction mixture, 20 ng genomic DNA or 2 μl 1:10-diluted cultured strains were used as the template and mixed with 0.4 μM RF-specific or IF-specific primers and a polymerase mixture from the Qiagen HotStarTaq Master Mix Kit using a three-step cycling program consisting of initial denaturation at 95°C for 15 min; 30 cycles of a three-step procedure including denaturation at 94°C for 30s, annealing at 52°C for 30s, and extension at 72°C for 30s; and a final extension at 72°C for 5 min. .. The PCR products were separated on a 1.8% agarose gel prepared with 0.5× Tris-borate-EDTA buffer.

    Hot Start PCR:

    Article Title: Site-specific methylation of placental HSD11B2 gene promoter is related to intrauterine growth restriction
    Article Snippet: .. Hot-start PCR was carried out with HotStar Taq Master Mix Kit (Qiagen) and each PCR mix consisted of the converted DNA, 1.5 m M MgCl2 , 200 μ M dNTPs, 10 pmol forward primer, 10 pmol reverse primer, and 1.25 U HotStar Taq Polymerase. .. The presence or absence of PCR products and freedom from PCR contamination was established on 1.5% agarose gels with ethidium bromide staining.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The Inactive 44-kDa Processed Form of Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Enhances Proteolytic Activity via Regulation of Endocytosis of Active MT1-MMP *The Inactive 44-kDa Processed Form of Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Enhances Proteolytic Activity via Regulation of Endocytosis of Active MT1-MMP * S⃞
    Article Snippet: .. RT-PCR was performed with 1 μg of each total RNA sample using SuperScript™ III reverse transcriptase (Invitrogen) and subsequently HotStar Taq® master mix kit (Qiagen) following the manufacturer's instructions. .. The sequences of the specific primers (IDT, Coralville, IA) for human TIMP-2 used are as follows: forward, 5′-GAA ACG ACA TTT ATG GCA ACC-3′; reverse, 5′-GCT GGA CCA GTC GAA ACC-3′.

    Expressing:

    Article Title: Use of Recombinant Entamoeba histolytica Cysteine Proteinase 1 To Identify a Potent Inhibitor of Amebic Invasion in a Human Colonic Model ▿
    Article Snippet: .. The sequence encoding the pro- and mature region of the E. histolytica EhCP1 gene (accession no. ) was amplified from E. histolytica genomic DNA by PCR (HotstartTaq Master Mix kit; QIAGEN, Valencia, CA) and cloned into the pBAD/Thio-TOPO plasmid (Invitrogen, Carlsbad, CA), resulting in expression of the recombinant protein as a thioredoxin fusion (amino terminus) with a six-residue histidine tail (carboxy terminus). .. Expression was induced as previously described , except that 0.2% arabinose was used for 4 h. For antibody production, rEhCP1 was expressed without the thioredoxin tail in the pRSETA vector (Invitrogen, Carlsbad, CA) and purified under denaturing conditions over nickel-nitrilotriacetic acid ( ).

    Sequencing:

    Article Title: Use of Recombinant Entamoeba histolytica Cysteine Proteinase 1 To Identify a Potent Inhibitor of Amebic Invasion in a Human Colonic Model ▿
    Article Snippet: .. The sequence encoding the pro- and mature region of the E. histolytica EhCP1 gene (accession no. ) was amplified from E. histolytica genomic DNA by PCR (HotstartTaq Master Mix kit; QIAGEN, Valencia, CA) and cloned into the pBAD/Thio-TOPO plasmid (Invitrogen, Carlsbad, CA), resulting in expression of the recombinant protein as a thioredoxin fusion (amino terminus) with a six-residue histidine tail (carboxy terminus). .. Expression was induced as previously described , except that 0.2% arabinose was used for 4 h. For antibody production, rEhCP1 was expressed without the thioredoxin tail in the pRSETA vector (Invitrogen, Carlsbad, CA) and purified under denaturing conditions over nickel-nitrilotriacetic acid ( ).

    Recombinant:

    Article Title: Use of Recombinant Entamoeba histolytica Cysteine Proteinase 1 To Identify a Potent Inhibitor of Amebic Invasion in a Human Colonic Model ▿
    Article Snippet: .. The sequence encoding the pro- and mature region of the E. histolytica EhCP1 gene (accession no. ) was amplified from E. histolytica genomic DNA by PCR (HotstartTaq Master Mix kit; QIAGEN, Valencia, CA) and cloned into the pBAD/Thio-TOPO plasmid (Invitrogen, Carlsbad, CA), resulting in expression of the recombinant protein as a thioredoxin fusion (amino terminus) with a six-residue histidine tail (carboxy terminus). .. Expression was induced as previously described , except that 0.2% arabinose was used for 4 h. For antibody production, rEhCP1 was expressed without the thioredoxin tail in the pRSETA vector (Invitrogen, Carlsbad, CA) and purified under denaturing conditions over nickel-nitrilotriacetic acid ( ).

    Plasmid Preparation:

    Article Title: Use of Recombinant Entamoeba histolytica Cysteine Proteinase 1 To Identify a Potent Inhibitor of Amebic Invasion in a Human Colonic Model ▿
    Article Snippet: .. The sequence encoding the pro- and mature region of the E. histolytica EhCP1 gene (accession no. ) was amplified from E. histolytica genomic DNA by PCR (HotstartTaq Master Mix kit; QIAGEN, Valencia, CA) and cloned into the pBAD/Thio-TOPO plasmid (Invitrogen, Carlsbad, CA), resulting in expression of the recombinant protein as a thioredoxin fusion (amino terminus) with a six-residue histidine tail (carboxy terminus). .. Expression was induced as previously described , except that 0.2% arabinose was used for 4 h. For antibody production, rEhCP1 was expressed without the thioredoxin tail in the pRSETA vector (Invitrogen, Carlsbad, CA) and purified under denaturing conditions over nickel-nitrilotriacetic acid ( ).

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    Qiagen hotstartaq master mix kit
    Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and <t>HotStarTaq</t> was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.
    Hotstartaq Master Mix Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 965 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.

    Journal: Experimental Animals

    Article Title: Zygosity Determination in Hairless Mice by PCR Based onHrhr Gene Analysis

    doi: 10.1538/expanim.62.267

    Figure Lengend Snippet: Analysis of Hr alleles in homozygous (H) and wild-type (W) HR mice. The normal (wild-type) Hr allele is ~19 kbp in length and consists of 19 exons. A total of 15 overlapping PCRs covering the entire Hr coding sequence revealed that an insertion mutation was localized in intron 6 of the Hr x allele. KOD FX neo was used for PCR amplification of regions 2–7, and HotStarTaq was used for PCR of other regions. PCRs shown in gray typeface (1, 2, 4, 6, 9, 12, 14, 15), no difference between homozygous and wild-type HR mice. PCRs shown in black typeface (3, 5, 7, 8, 10, 11, 13), no band or different bands were obtained in homozygous HR mice. Six agarose gel electropherograms show the band patterns of all PCRs. The primer sets used were: (1) F224 and R1151, (2) F193 and R1873, (3) F193 and R2433, (4) F2463 and R3455, (5) F1843 and R2433, (6) F2052 and R2433, (7) F1843 and R2078, (8) F1843 and R1972, (9) F1913 and R2078, (10) F1843 and Int6-R1458, (11) F1843 and Int6-R979, (12) Int6-F1290 and R1972, (13) F1843 and Int6-R850, (14) F1843 and Int6-R642, and (15) F1843 and Int6-R539. The primer sequences and elongation time are shown in Tables 1 and 2 , respectively. The primer positions for long PCR shown in Fig. 2 are also indicated in this figure.

    Article Snippet: The difference between homozygous (Hrx /Hrx ) and wild-type (Hr /Hr ) genomes was determined using multiple PCRs with HotStarTaq (#203443, Qiagen; regions 1, 8–13) or KOD FX neo (KFX-201, TOYOBO, Osaka, Japan; regions 2–7) DNA polymerases.

    Techniques: Mouse Assay, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis