type it hrm pcr kit  (Qiagen)


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    Name:
    Type it HRM PCR Kit
    Description:
    For fast and accurate detection of gene mutations and SNPs by High Resolution Melting HRM analysis Kit contents Qiagen Type it HRM PCR Kit 100 x 25μL rxns Genomic DNA Sample PCR amplification Reaction Type 5 3 Exonuclease Enzyme Activity For Fast and Accurate Detection of Gene Mutations and SNPs by High Resolution Melting HRM Analysis Includes 1 x 1 3mL of 2x HRM PCR Master Mix Contains HotStarTaq Plus DNA Polymerase EvaGreen Dye optimized concentration of Q solution dNTPs and MgCl2 and RNase free Water Benefits Reliable and accurate detection of subtle sequence variations Highly suited for use with any cycler with HRM capabilities Distinct melting curves due to novel EvaGreen fluorescent dye Easy development of reliable new HRM genotyping assays Convenient master mix format and optimized protocols
    Catalog Number:
    206542
    Price:
    99.3
    Category:
    Type it HRM PCR Kit
    Buy from Supplier


    Structured Review

    Qiagen type it hrm pcr kit
    Type it HRM PCR Kit
    For fast and accurate detection of gene mutations and SNPs by High Resolution Melting HRM analysis Kit contents Qiagen Type it HRM PCR Kit 100 x 25μL rxns Genomic DNA Sample PCR amplification Reaction Type 5 3 Exonuclease Enzyme Activity For Fast and Accurate Detection of Gene Mutations and SNPs by High Resolution Melting HRM Analysis Includes 1 x 1 3mL of 2x HRM PCR Master Mix Contains HotStarTaq Plus DNA Polymerase EvaGreen Dye optimized concentration of Q solution dNTPs and MgCl2 and RNase free Water Benefits Reliable and accurate detection of subtle sequence variations Highly suited for use with any cycler with HRM capabilities Distinct melting curves due to novel EvaGreen fluorescent dye Easy development of reliable new HRM genotyping assays Convenient master mix format and optimized protocols
    https://www.bioz.com/result/type it hrm pcr kit/product/Qiagen
    Average 99 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    type it hrm pcr kit - by Bioz Stars, 2020-03
    99/100 stars

    Images

    1) Product Images from "Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis"

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms151119898

    The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.
    Figure Legend Snippet: The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.

    Techniques Used: Real-time Polymerase Chain Reaction

    The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.
    Figure Legend Snippet: The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.

    Techniques Used: Real-time Polymerase Chain Reaction

    2) Product Images from "Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis"

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis

    Journal: F1000Research

    doi: 10.12688/f1000research.2-281.v2

    Detection of semen in reportedly single source vaginal samples using a capillary electrophoresis multiple mRNA profiling assay. RT-PCR products from RNA extracted from previously reported single source vaginal secretions samples were amplified in a multiplex reaction using a mRNA profiling assay for body fluid/tissue identification. The multiplex contains mRNA biomarkers for blood, semen, saliva, vaginal secretions, menstrual blood and skin. Shown here are two vaginal samples ( A and B ) in which semen was detected as indicated by the presence of the PRM2 and TGM4 (semen-specific biomarkers). This data is consistent with the detection of semen using the semen-saliva duplex HRM assay. The presence of vaginal secretions is indicated by the presence of CYP2B7P1. The x-axis indicates size in base pairs and the y-axis indicates relative fluorescence units (note: the y-axis scales are different for some panels due to varying signal intensities of the observed products). The grey areas represent the bins for biomarker identification. The biomarker name and relative fluorescence units are displayed.
    Figure Legend Snippet: Detection of semen in reportedly single source vaginal samples using a capillary electrophoresis multiple mRNA profiling assay. RT-PCR products from RNA extracted from previously reported single source vaginal secretions samples were amplified in a multiplex reaction using a mRNA profiling assay for body fluid/tissue identification. The multiplex contains mRNA biomarkers for blood, semen, saliva, vaginal secretions, menstrual blood and skin. Shown here are two vaginal samples ( A and B ) in which semen was detected as indicated by the presence of the PRM2 and TGM4 (semen-specific biomarkers). This data is consistent with the detection of semen using the semen-saliva duplex HRM assay. The presence of vaginal secretions is indicated by the presence of CYP2B7P1. The x-axis indicates size in base pairs and the y-axis indicates relative fluorescence units (note: the y-axis scales are different for some panels due to varying signal intensities of the observed products). The grey areas represent the bins for biomarker identification. The biomarker name and relative fluorescence units are displayed.

    Techniques Used: Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Amplification, Multiplex Assay, HRM Assay, Fluorescence, Biomarker Assay

    3) Product Images from "Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis"

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms151119898

    The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.
    Figure Legend Snippet: The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.

    Techniques Used: Real-time Polymerase Chain Reaction

    The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.
    Figure Legend Snippet: The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.

    Techniques Used: Real-time Polymerase Chain Reaction

    4) Product Images from "Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis"

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis

    Journal: F1000Research

    doi: 10.12688/f1000research.2-281.v2

    Detection of semen in reportedly single source vaginal samples using a capillary electrophoresis multiple mRNA profiling assay. RT-PCR products from RNA extracted from previously reported single source vaginal secretions samples were amplified in a multiplex reaction using a mRNA profiling assay for body fluid/tissue identification. The multiplex contains mRNA biomarkers for blood, semen, saliva, vaginal secretions, menstrual blood and skin. Shown here are two vaginal samples ( A and B ) in which semen was detected as indicated by the presence of the PRM2 and TGM4 (semen-specific biomarkers). This data is consistent with the detection of semen using the semen-saliva duplex HRM assay. The presence of vaginal secretions is indicated by the presence of CYP2B7P1. The x-axis indicates size in base pairs and the y-axis indicates relative fluorescence units (note: the y-axis scales are different for some panels due to varying signal intensities of the observed products). The grey areas represent the bins for biomarker identification. The biomarker name and relative fluorescence units are displayed.
    Figure Legend Snippet: Detection of semen in reportedly single source vaginal samples using a capillary electrophoresis multiple mRNA profiling assay. RT-PCR products from RNA extracted from previously reported single source vaginal secretions samples were amplified in a multiplex reaction using a mRNA profiling assay for body fluid/tissue identification. The multiplex contains mRNA biomarkers for blood, semen, saliva, vaginal secretions, menstrual blood and skin. Shown here are two vaginal samples ( A and B ) in which semen was detected as indicated by the presence of the PRM2 and TGM4 (semen-specific biomarkers). This data is consistent with the detection of semen using the semen-saliva duplex HRM assay. The presence of vaginal secretions is indicated by the presence of CYP2B7P1. The x-axis indicates size in base pairs and the y-axis indicates relative fluorescence units (note: the y-axis scales are different for some panels due to varying signal intensities of the observed products). The grey areas represent the bins for biomarker identification. The biomarker name and relative fluorescence units are displayed.

    Techniques Used: Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Amplification, Multiplex Assay, HRM Assay, Fluorescence, Biomarker Assay

    Related Articles

    DNA Extraction:

    Article Title: Association between Ala379Val polymorphism of lipoprotein-associated phospholipase A2 and migraine without aura in Iranian population
    Article Snippet: Genomic DNA samples were extracted from peripheral whole blood using the AccuPrep Genomic DNA Extraction kit (Bioneer Inc., Korea) according to the manufacturer’s protocol. .. Polymerase chain reactions (PCRs) were carried out in duplicate in 20 µl of final volume using the type-it HRM kit (Qiagen), HRM PCR buffer, HotStarTaq Plus DNA Polymerase, nucleotides and EvaGreen dye, and 30 ng DNA.

    Article Title: Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology
    Article Snippet: Paragraph title: DNA Extraction and HRM-PCR Amplification ... Reaction mixtures had a final volume of 25 μl, and contained 50 ng genomic DNA, 12.5 μL of 2 × HRM PCR master mix (Type-it HRMTM PCR Kit, Qiagen), 1 μL of 10 μM forward and reverse primers, and distilled water was added up to the final volume.

    Clone Assay:

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
    Article Snippet: .. The Zero Blunt TOPO PCR Cloning kit (Invitrogen, Vienna, Austria), EasyPrep Pro Plasmid Miniprep kit (Biozym), JM109 Competent Cells ( > 107 cfu/μg, Promega, Mannheim, Germany), Wizard DNA Clean-up System (Promega), Phusion Hot Start II High-Fidelity DNA Polymerase kit (Biozym), and Type-it HRM PCR Kit (Qiagen, Hilden, Germany) were used according to the respective manufacturer’s recommendations. ..

    Multiplex Assay:

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR
    Article Snippet: .. Identification and quantification of ETEC fimbriae genes by individual / multiplex HRM-qPCR All HRM-qPCR reactions were performed using a Rotor-Gene Q (QIAGEN) HRM-thermo cycler and Type-it HRM Kit (QIAGEN) Primers targeting five different porcine ETEC fimbriae genes (K99, F41, F18, F6 and K88) were designed with nearly identical annealing temperature (62 °C to 63 °C) (Table ) to allow amplification in multiplex PCR reaction. .. HRM-qPCR reactions contained 12.5 μL 2× HRM Master Mix, 2 μL template DNA for individual reaction or 3 μL for multiplex reaction, 700 nM primers for individual reaction and 200 nM per target for multiplex detection to a final volume of 25 μL.

    Article Title: Hepatocytes from wild-type or heterozygous donors are equally effective in achieving successful therapeutic liver repopulation in murine phenylketonuria (PKU)
    Article Snippet: The reactions were catalyzed using standard conditions with HotStar Taq Plus™ DNA polymerase in the presence of EvaGreen™ fluorescent dye (Type-It HRM PCR kit™ , Qiagen Inc., Valencia, CA) in a Rotorgene Q™ real-time thermocycler with built in high resolution melt analysis capability according to the manufacturer’s instructions. .. This multiplex PCR reaction yielded amplicons of 85 bp and 64 bp from the wild-type and FahΔexon5 genomes respectively.

    Amplification:

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR
    Article Snippet: .. Identification and quantification of ETEC fimbriae genes by individual / multiplex HRM-qPCR All HRM-qPCR reactions were performed using a Rotor-Gene Q (QIAGEN) HRM-thermo cycler and Type-it HRM Kit (QIAGEN) Primers targeting five different porcine ETEC fimbriae genes (K99, F41, F18, F6 and K88) were designed with nearly identical annealing temperature (62 °C to 63 °C) (Table ) to allow amplification in multiplex PCR reaction. .. HRM-qPCR reactions contained 12.5 μL 2× HRM Master Mix, 2 μL template DNA for individual reaction or 3 μL for multiplex reaction, 700 nM primers for individual reaction and 200 nM per target for multiplex detection to a final volume of 25 μL.

    Article Title: Comparative-high resolution melting: a novel method of simultaneous screening for small mutations and copy number variations
    Article Snippet: .. Assay design The products were amplified using the type-it HRM kit (Qiagen) on the DNA templates at a concentration of 50 ng/μl diluted in AE buffer (Qiagen). .. The analysis was performed on a Rotor-Gene® Q equipment (Qiagen).

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures
    Article Snippet: PCR HRM Analysis Real-time PCR amplification and HRM analysis of ITS and 16S rRNA gene was performed on the Rotor-Gene Q thermal cycler (Qiagen, Venlo, Netherlands) with ITS primers pairs ITS1F (5′-TTGTACACACCGCCCG-3′) and ITS2R (5′-YGCCAAGGCATCCACC-3′) and 16S primers pairs V1F (5′-GYGGCGNACGGGTGAGTAA-3′) and V6R (5′-AGCTGACGACANCCATGCA-3′). .. PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA.

    Article Title: High Resolution Melting analysis as a rapid and efficient method of screening for small mutations in the STK11 gene in patients with Peutz-Jeghers syndrome
    Article Snippet: .. Fragments were amplified using Type-it HRM kit [Qiagen] and the analysis was performed on a Rotor-Gene Q equipment [Qiagen]. ..

    Article Title: Differential Identification of Mycobacterial Species Using High-Resolution Melting Analysis
    Article Snippet: Real-Time PCR-HRM Assay Real-time PCR was performed with a Type-it HRM PCR Kit (QIAGEN, Germany) on a Light Cycler 480 system (Roche Diagnostics, Switzerland). .. The amplification was performed using the following conditions: a pre-incubation step at 95°C for 10 min, followed by 45 cycles of denaturation at 95°C for 10 s, annealing at 65°C for 30 s, and extension at 72°C for 10 s followed by the Tm analysis with increasing temperatures from 60 to 95°C in a 0.2°C s-1 slope increment for 10 s. The HRM analysis was performed using Gene Scanning Software Version 1.5.0 (Roche Instrument Centre, Switzerland).

    Article Title: A Rare Missense Mutation and a Polymorphism with High Frequency in LDLR Gene among Iranian Patients with Familial Hypercholesterolemia
    Article Snippet: PCR and high-resolution melting (HRM) were accomplished by Rotor-Gene Q 2plx HRM (Qiagen, Hilden, Germany) using Type-it HRM PCR Kit (Qiagen, Hilden, Germany) with 60 ng of gDNA and 13 pmol of each primer in a final volume of 25 μl, in accordance with manufacturer's instruction. .. The amplified segment of exon 9, including coding region and exon-intron boundaries of LDLR , was screened by HRM as described in Whittall et al . procedure[ ] and the samples with a shift in HRM and melting temperature curves were then sequenced.

    Article Title: Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology
    Article Snippet: Paragraph title: DNA Extraction and HRM-PCR Amplification ... Reaction mixtures had a final volume of 25 μl, and contained 50 ng genomic DNA, 12.5 μL of 2 × HRM PCR master mix (Type-it HRMTM PCR Kit, Qiagen), 1 μL of 10 μM forward and reverse primers, and distilled water was added up to the final volume.

    Article Title: Rapid Identification of Officinal Akebiae Caulis and Its Toxic Adulterant Aristolochiae Manshuriensis Caulis (Aristolochia manshuriensis) by Loop-Mediated Isothermal Amplification
    Article Snippet: We implemented the PCR reaction at 94°C for 5 min; followed by 40 cycles at 94°C for 30 s, 56°C for 30 s, and 72°C for 45 s; and a final extension step at 72°C for 10 min. We inspected the amplification products by 1% agarose gel electrophoresis. .. Furthermore, real-time PCR was carried out in Rotor-Gene Q (QIAGEN, Germany) by using 12.5 μL 2 × HRM PCR master mix (Type-it® HRMTM PCR Kit, QIAGEN, Germany).

    Article Title: Hepatocytes from wild-type or heterozygous donors are equally effective in achieving successful therapeutic liver repopulation in murine phenylketonuria (PKU)
    Article Snippet: In this assay, 100 ng hepatocyte genomic DNA isolated by standard proteinase digestion and phenol/chloroform extraction was subjected to PCR amplification of the FAH gene in the region spanning the 5′ junction of the FahΔexon5 insertion ( ). .. The reactions were catalyzed using standard conditions with HotStar Taq Plus™ DNA polymerase in the presence of EvaGreen™ fluorescent dye (Type-It HRM PCR kit™ , Qiagen Inc., Valencia, CA) in a Rotorgene Q™ real-time thermocycler with built in high resolution melt analysis capability according to the manufacturer’s instructions.

    Agarose Gel Electrophoresis:

    Article Title: High Resolution Melting analysis as a rapid and efficient method of screening for small mutations in the STK11 gene in patients with Peutz-Jeghers syndrome
    Article Snippet: In addition, PCR product heterogeneity on agarose gel (1.5% agarose gel, electrophoresis – 30 min., voltage – 100 V) was also checked. .. Fragments were amplified using Type-it HRM kit [Qiagen] and the analysis was performed on a Rotor-Gene Q equipment [Qiagen].

    Article Title: Rapid Identification of Officinal Akebiae Caulis and Its Toxic Adulterant Aristolochiae Manshuriensis Caulis (Aristolochia manshuriensis) by Loop-Mediated Isothermal Amplification
    Article Snippet: We implemented the PCR reaction at 94°C for 5 min; followed by 40 cycles at 94°C for 30 s, 56°C for 30 s, and 72°C for 45 s; and a final extension step at 72°C for 10 min. We inspected the amplification products by 1% agarose gel electrophoresis. .. Furthermore, real-time PCR was carried out in Rotor-Gene Q (QIAGEN, Germany) by using 12.5 μL 2 × HRM PCR master mix (Type-it® HRMTM PCR Kit, QIAGEN, Germany).

    Synthesized:

    Article Title: Association between Ala379Val polymorphism of lipoprotein-associated phospholipase A2 and migraine without aura in Iranian population
    Article Snippet: The single-nucleotide polymorphisms (SNPs) rs1051931 (A379V) were identified by the National Center for Biotechnology Information (NCBI) data bank and primers were designed by Beacon Designer 8.00 to flank the coding regions (PREMIER Biosoft International, USA and synthesized by TIB MOLBIOL, Germany). .. Polymerase chain reactions (PCRs) were carried out in duplicate in 20 µl of final volume using the type-it HRM kit (Qiagen), HRM PCR buffer, HotStarTaq Plus DNA Polymerase, nucleotides and EvaGreen dye, and 30 ng DNA.

    Isolation:

    Article Title: Hepatocytes from wild-type or heterozygous donors are equally effective in achieving successful therapeutic liver repopulation in murine phenylketonuria (PKU)
    Article Snippet: In this assay, 100 ng hepatocyte genomic DNA isolated by standard proteinase digestion and phenol/chloroform extraction was subjected to PCR amplification of the FAH gene in the region spanning the 5′ junction of the FahΔexon5 insertion ( ). .. The reactions were catalyzed using standard conditions with HotStar Taq Plus™ DNA polymerase in the presence of EvaGreen™ fluorescent dye (Type-It HRM PCR kit™ , Qiagen Inc., Valencia, CA) in a Rotorgene Q™ real-time thermocycler with built in high resolution melt analysis capability according to the manufacturer’s instructions.

    HRM Assay:

    Article Title: Association between Ala379Val polymorphism of lipoprotein-associated phospholipase A2 and migraine without aura in Iranian population
    Article Snippet: Genotyping was done by high-resolution melt (HRM) assay using a Rotor-Gene 6000 instrument (Corbett Life Science, Australia). .. Polymerase chain reactions (PCRs) were carried out in duplicate in 20 µl of final volume using the type-it HRM kit (Qiagen), HRM PCR buffer, HotStarTaq Plus DNA Polymerase, nucleotides and EvaGreen dye, and 30 ng DNA.

    Size-exclusion Chromatography:

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis
    Article Snippet: High Resolution Melt (HRM) analysis Singleplex, Duplex, Triplex Assays: HRM assays were performed using the Type-It® HRM™ PCR kit (QIAGEN, Germantown, MD). .. All assays were run on the Rotor-Gene® Q real time PCR instrument (QIAGEN), using the following cycling conditions: 95°C 5 min, followed by 45 cycles of 94°C 10 sec, 55°C 30 sec, 72°C 10 sec. HRM analysis was performed using a 65–90°C temperature range, with +0.1°C increments (90 sec of pre-melt conditioning on first step; and 2 sec for each step afterwards).

    Article Title: Genome sequencing and analysis of Mangalica, a fatty local pig of Hungary
    Article Snippet: PCR reactions were performed in 25 μl reaction volumes using 60 ng total DNA as template and the Type-it HRM PCR kit (Qiagen, Hilden, Germany), according to the instruction of the manufacturer. .. Reactions were carried out with an initial denaturation step at 95°C for 5 min, followed by 35 cycles of 95°C for 15 sec, 60°C for 30 sec and 72°C for 10 sec and then HRM curves were generated by acquiring florescence data between 80 and 91°C.

    Real-time Polymerase Chain Reaction:

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis
    Article Snippet: High Resolution Melt (HRM) analysis Singleplex, Duplex, Triplex Assays: HRM assays were performed using the Type-It® HRM™ PCR kit (QIAGEN, Germantown, MD). .. All assays were run on the Rotor-Gene® Q real time PCR instrument (QIAGEN), using the following cycling conditions: 95°C 5 min, followed by 45 cycles of 94°C 10 sec, 55°C 30 sec, 72°C 10 sec. HRM analysis was performed using a 65–90°C temperature range, with +0.1°C increments (90 sec of pre-melt conditioning on first step; and 2 sec for each step afterwards).

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures
    Article Snippet: PCR HRM Analysis Real-time PCR amplification and HRM analysis of ITS and 16S rRNA gene was performed on the Rotor-Gene Q thermal cycler (Qiagen, Venlo, Netherlands) with ITS primers pairs ITS1F (5′-TTGTACACACCGCCCG-3′) and ITS2R (5′-YGCCAAGGCATCCACC-3′) and 16S primers pairs V1F (5′-GYGGCGNACGGGTGAGTAA-3′) and V6R (5′-AGCTGACGACANCCATGCA-3′). .. PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA.

    Article Title: Differential Identification of Mycobacterial Species Using High-Resolution Melting Analysis
    Article Snippet: .. Real-Time PCR-HRM Assay Real-time PCR was performed with a Type-it HRM PCR Kit (QIAGEN, Germany) on a Light Cycler 480 system (Roche Diagnostics, Switzerland). .. Each PCR analysis contained one primer pair.

    Article Title: Rapid Identification of Officinal Akebiae Caulis and Its Toxic Adulterant Aristolochiae Manshuriensis Caulis (Aristolochia manshuriensis) by Loop-Mediated Isothermal Amplification
    Article Snippet: .. Furthermore, real-time PCR was carried out in Rotor-Gene Q (QIAGEN, Germany) by using 12.5 μL 2 × HRM PCR master mix (Type-it® HRMTM PCR Kit, QIAGEN, Germany). ..

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
    Article Snippet: Paragraph title: 3.5. Real-Time PCR and HRM Conditions ... The Type-it HRM PCR Kit (Qiagen), 700 nM primers (Scorpion primer 125 nM), and 1.6 µL of undiluted DNA (70–100 ng/µL) template were used for each reaction.

    Concentration Assay:

    Article Title: Comparative-high resolution melting: a novel method of simultaneous screening for small mutations and copy number variations
    Article Snippet: .. Assay design The products were amplified using the type-it HRM kit (Qiagen) on the DNA templates at a concentration of 50 ng/μl diluted in AE buffer (Qiagen). .. The analysis was performed on a Rotor-Gene® Q equipment (Qiagen).

    Incubation:

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures
    Article Snippet: PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA. .. Before adding the low temperature calibrator and genomic DNA, each reaction was treated with 0.5 μL of dsDNase and 0.5 μL DTT (ArcticZymes PCR Decontamination Kit, ArcticZymes, Tromsø, Norway) followed by incubation at 37 °C for 20 minutes and 60 °C for 20 minutes to eliminate possible contaminating DNA.

    Polymerase Chain Reaction:

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis
    Article Snippet: .. High Resolution Melt (HRM) analysis Singleplex, Duplex, Triplex Assays: HRM assays were performed using the Type-It® HRM™ PCR kit (QIAGEN, Germantown, MD). ..

    Article Title: Association between Ala379Val polymorphism of lipoprotein-associated phospholipase A2 and migraine without aura in Iranian population
    Article Snippet: .. Polymerase chain reactions (PCRs) were carried out in duplicate in 20 µl of final volume using the type-it HRM kit (Qiagen), HRM PCR buffer, HotStarTaq Plus DNA Polymerase, nucleotides and EvaGreen dye, and 30 ng DNA. .. The PCR program consisted of an initial denaturation-activation step at 95 °C for 5 minutes, followed by a 40-cycle program (denaturation at 95 °C for 15 seconds, annealing conditions 55 °C for 5 seconds, 72 °C for 15 seconds; a HRM step from 70 to 95 °C rising at 0.1 °C per second).

    Article Title: Genome sequencing and analysis of Mangalica, a fatty local pig of Hungary
    Article Snippet: .. PCR reactions were performed in 25 μl reaction volumes using 60 ng total DNA as template and the Type-it HRM PCR kit (Qiagen, Hilden, Germany), according to the instruction of the manufacturer. ..

    Article Title: Comparative-high resolution melting: a novel method of simultaneous screening for small mutations and copy number variations
    Article Snippet: Assay design The products were amplified using the type-it HRM kit (Qiagen) on the DNA templates at a concentration of 50 ng/μl diluted in AE buffer (Qiagen). .. PCR reactions were carried out for the 30 cycles (with a 5 min preincubation at 95 °C) of 95 °C for 10 s, 55 °C for 30 s and 72 °C for 10 s, the products were then melted and PCR was continued to the 40th cycle in the same conditions followed by another melting process.

    Article Title: Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis
    Article Snippet: .. ‘All Body Fluids’ Hexaplex Assay: HRM assays were performed using the Type-It® HRM™ PCR kit (QIAGEN, Germantown, MD). ..

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures
    Article Snippet: .. PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA. .. Before adding the low temperature calibrator and genomic DNA, each reaction was treated with 0.5 μL of dsDNase and 0.5 μL DTT (ArcticZymes PCR Decontamination Kit, ArcticZymes, Tromsø, Norway) followed by incubation at 37 °C for 20 minutes and 60 °C for 20 minutes to eliminate possible contaminating DNA.

    Article Title: High Resolution Melting analysis as a rapid and efficient method of screening for small mutations in the STK11 gene in patients with Peutz-Jeghers syndrome
    Article Snippet: The obtained PCR amplicons were contained within the length range of 150–250 bp, while primers annealing temperature was 60°C for all pairs. .. Fragments were amplified using Type-it HRM kit [Qiagen] and the analysis was performed on a Rotor-Gene Q equipment [Qiagen].

    Article Title: Differential Identification of Mycobacterial Species Using High-Resolution Melting Analysis
    Article Snippet: .. Real-Time PCR-HRM Assay Real-time PCR was performed with a Type-it HRM PCR Kit (QIAGEN, Germany) on a Light Cycler 480 system (Roche Diagnostics, Switzerland). .. Each PCR analysis contained one primer pair.

    Article Title: A Rare Missense Mutation and a Polymorphism with High Frequency in LDLR Gene among Iranian Patients with Familial Hypercholesterolemia
    Article Snippet: .. PCR and high-resolution melting (HRM) were accomplished by Rotor-Gene Q 2plx HRM (Qiagen, Hilden, Germany) using Type-it HRM PCR Kit (Qiagen, Hilden, Germany) with 60 ng of gDNA and 13 pmol of each primer in a final volume of 25 μl, in accordance with manufacturer's instruction. .. The amplified segment of exon 9, including coding region and exon-intron boundaries of LDLR , was screened by HRM as described in Whittall et al . procedure[ ] and the samples with a shift in HRM and melting temperature curves were then sequenced.

    Article Title: Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology
    Article Snippet: .. Reaction mixtures had a final volume of 25 μl, and contained 50 ng genomic DNA, 12.5 μL of 2 × HRM PCR master mix (Type-it HRMTM PCR Kit, Qiagen), 1 μL of 10 μM forward and reverse primers, and distilled water was added up to the final volume. ..

    Article Title: Rapid Identification of Officinal Akebiae Caulis and Its Toxic Adulterant Aristolochiae Manshuriensis Caulis (Aristolochia manshuriensis) by Loop-Mediated Isothermal Amplification
    Article Snippet: .. Furthermore, real-time PCR was carried out in Rotor-Gene Q (QIAGEN, Germany) by using 12.5 μL 2 × HRM PCR master mix (Type-it® HRMTM PCR Kit, QIAGEN, Germany). ..

    Article Title: Hepatocytes from wild-type or heterozygous donors are equally effective in achieving successful therapeutic liver repopulation in murine phenylketonuria (PKU)
    Article Snippet: .. The reactions were catalyzed using standard conditions with HotStar Taq Plus™ DNA polymerase in the presence of EvaGreen™ fluorescent dye (Type-It HRM PCR kit™ , Qiagen Inc., Valencia, CA) in a Rotorgene Q™ real-time thermocycler with built in high resolution melt analysis capability according to the manufacturer’s instructions. ..

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
    Article Snippet: .. The Zero Blunt TOPO PCR Cloning kit (Invitrogen, Vienna, Austria), EasyPrep Pro Plasmid Miniprep kit (Biozym), JM109 Competent Cells ( > 107 cfu/μg, Promega, Mannheim, Germany), Wizard DNA Clean-up System (Promega), Phusion Hot Start II High-Fidelity DNA Polymerase kit (Biozym), and Type-it HRM PCR Kit (Qiagen, Hilden, Germany) were used according to the respective manufacturer’s recommendations. ..

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
    Article Snippet: .. The Type-it HRM PCR Kit (Qiagen), 700 nM primers (Scorpion primer 125 nM), and 1.6 µL of undiluted DNA (70–100 ng/µL) template were used for each reaction. ..

    Sequencing:

    Article Title: Association between Ala379Val polymorphism of lipoprotein-associated phospholipase A2 and migraine without aura in Iranian population
    Article Snippet: Polymerase chain reactions (PCRs) were carried out in duplicate in 20 µl of final volume using the type-it HRM kit (Qiagen), HRM PCR buffer, HotStarTaq Plus DNA Polymerase, nucleotides and EvaGreen dye, and 30 ng DNA. .. Normalized and temperature-shifted melting curves from HRM, suggestive of SNPs, were distinguished and the samples were subjected to direct sequencing.

    Article Title: Comparative-high resolution melting: a novel method of simultaneous screening for small mutations and copy number variations
    Article Snippet: Assay design The products were amplified using the type-it HRM kit (Qiagen) on the DNA templates at a concentration of 50 ng/μl diluted in AE buffer (Qiagen). .. The first melting analysis was performed from 70 °C to 90 °C by raising the temperature by 0.3° at each step after which the second one, designed to detect small changes in the sequence, was carried out with higher resolution raising the temperature by 0.1° at each step.

    Software:

    Article Title: Genome sequencing and analysis of Mangalica, a fatty local pig of Hungary
    Article Snippet: PCR reactions were performed in 25 μl reaction volumes using 60 ng total DNA as template and the Type-it HRM PCR kit (Qiagen, Hilden, Germany), according to the instruction of the manufacturer. .. Individuals with homozygous and heterozygous genotypes were assigned according to their HRM curve determined by the Rotor-Gene software and visual inspection.

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures
    Article Snippet: PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA. .. HRM analysis was performed utilizing the RotorGene-Q software.

    Article Title: Differential Identification of Mycobacterial Species Using High-Resolution Melting Analysis
    Article Snippet: Real-Time PCR-HRM Assay Real-time PCR was performed with a Type-it HRM PCR Kit (QIAGEN, Germany) on a Light Cycler 480 system (Roche Diagnostics, Switzerland). .. The amplification was performed using the following conditions: a pre-incubation step at 95°C for 10 min, followed by 45 cycles of denaturation at 95°C for 10 s, annealing at 65°C for 30 s, and extension at 72°C for 10 s followed by the Tm analysis with increasing temperatures from 60 to 95°C in a 0.2°C s-1 slope increment for 10 s. The HRM analysis was performed using Gene Scanning Software Version 1.5.0 (Roche Instrument Centre, Switzerland).

    Plasmid Preparation:

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
    Article Snippet: .. The Zero Blunt TOPO PCR Cloning kit (Invitrogen, Vienna, Austria), EasyPrep Pro Plasmid Miniprep kit (Biozym), JM109 Competent Cells ( > 107 cfu/μg, Promega, Mannheim, Germany), Wizard DNA Clean-up System (Promega), Phusion Hot Start II High-Fidelity DNA Polymerase kit (Biozym), and Type-it HRM PCR Kit (Qiagen, Hilden, Germany) were used according to the respective manufacturer’s recommendations. ..

    Generated:

    Article Title: Genome sequencing and analysis of Mangalica, a fatty local pig of Hungary
    Article Snippet: PCR reactions were performed in 25 μl reaction volumes using 60 ng total DNA as template and the Type-it HRM PCR kit (Qiagen, Hilden, Germany), according to the instruction of the manufacturer. .. Reactions were carried out with an initial denaturation step at 95°C for 5 min, followed by 35 cycles of 95°C for 15 sec, 60°C for 30 sec and 72°C for 10 sec and then HRM curves were generated by acquiring florescence data between 80 and 91°C.

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    Qiagen type it hrm kit
    Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by <t>qPCR</t> and multiplex <t>HRM-qPCR</t> methods ( n = 14)
    Type It Hrm Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/type it hrm kit/product/Qiagen
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    Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)

    Journal: BMC Microbiology

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR

    doi: 10.1186/s12866-017-1023-5

    Figure Lengend Snippet: Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)

    Article Snippet: Identification and quantification of ETEC fimbriae genes by individual / multiplex HRM-qPCR All HRM-qPCR reactions were performed using a Rotor-Gene Q (QIAGEN) HRM-thermo cycler and Type-it HRM Kit (QIAGEN) Primers targeting five different porcine ETEC fimbriae genes (K99, F41, F18, F6 and K88) were designed with nearly identical annealing temperature (62 °C to 63 °C) (Table ) to allow amplification in multiplex PCR reaction.

    Techniques: Real-time Polymerase Chain Reaction, Multiplex Assay

    Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )

    Journal: BMC Microbiology

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR

    doi: 10.1186/s12866-017-1023-5

    Figure Lengend Snippet: Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )

    Article Snippet: Identification and quantification of ETEC fimbriae genes by individual / multiplex HRM-qPCR All HRM-qPCR reactions were performed using a Rotor-Gene Q (QIAGEN) HRM-thermo cycler and Type-it HRM Kit (QIAGEN) Primers targeting five different porcine ETEC fimbriae genes (K99, F41, F18, F6 and K88) were designed with nearly identical annealing temperature (62 °C to 63 °C) (Table ) to allow amplification in multiplex PCR reaction.

    Techniques: Sequencing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Positive Control, Multiplex Assay, Software

    Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.

    Journal: Scientific Reports

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures

    doi: 10.1038/srep42097

    Figure Lengend Snippet: Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.

    Article Snippet: PCR reactions were performed in a 20 μL volume using Type-It HRM kit (Qiagen, Venlo, Netherlands) with the addition of 200 nM low temperature calibrator and 2 μL genomic DNA.

    Techniques: Sequencing, Polymerase Chain Reaction, Marker, Agarose Gel Electrophoresis, Molecular Weight

    The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.

    Journal: International Journal of Molecular Sciences

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

    doi: 10.3390/ijms151119898

    Figure Lengend Snippet: The distribution of the HRM measurements of the 5' junction of MON810 (5'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted on the x -axis and the numbers of the samples for the classes on the y -axis.

    Article Snippet: The Type-it HRM PCR Kit (Qiagen), 700 nM primers (Scorpion primer 125 nM), and 1.6 µL of undiluted DNA (70–100 ng/µL) template were used for each reaction.

    Techniques: Real-time Polymerase Chain Reaction

    The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.

    Journal: International Journal of Molecular Sciences

    Article Title: Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

    doi: 10.3390/ijms151119898

    Figure Lengend Snippet: The distribution of the HRM measurements of the 3' junction of MON810 (3'-MON810) region tested with real-time PCR. For the HRM analysis the confidence values were divided into 10 classes. The values of these classes were plotted against the number of samples.

    Article Snippet: The Type-it HRM PCR Kit (Qiagen), 700 nM primers (Scorpion primer 125 nM), and 1.6 µL of undiluted DNA (70–100 ng/µL) template were used for each reaction.

    Techniques: Real-time Polymerase Chain Reaction

    Sensitivity of C-HRM in CNV detection. The sensitivity of the C-HRM method was assessed by mixing different proportions of the DNA from female deletion carrier of the DMD exon 45 and wild type control ( a ). The results are presented as a linear regression with R 2 of 0.9974 ( b ). Taking into consideration the scale we adopted, theoretically it could be possible to distinguish a sample with only 17.3 % of DNA with the deletion

    Journal: Human Genetics

    Article Title: Comparative-high resolution melting: a novel method of simultaneous screening for small mutations and copy number variations

    doi: 10.1007/s00439-013-1393-1

    Figure Lengend Snippet: Sensitivity of C-HRM in CNV detection. The sensitivity of the C-HRM method was assessed by mixing different proportions of the DNA from female deletion carrier of the DMD exon 45 and wild type control ( a ). The results are presented as a linear regression with R 2 of 0.9974 ( b ). Taking into consideration the scale we adopted, theoretically it could be possible to distinguish a sample with only 17.3 % of DNA with the deletion

    Article Snippet: Assay design The products were amplified using the type-it HRM kit (Qiagen) on the DNA templates at a concentration of 50 ng/μl diluted in AE buffer (Qiagen).

    Techniques: