longamp taq  (New England Biolabs)


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    Name:
    LongAmp Taq 2X Master Mix
    Description:
    LongAmp Taq 2X Master Mix 500 rxns
    Catalog Number:
    M0287L
    Price:
    537
    Category:
    Long distance PCR Kits
    Size:
    500 rxns
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    Structured Review

    New England Biolabs longamp taq
    LongAmp Taq 2X Master Mix
    LongAmp Taq 2X Master Mix 500 rxns
    https://www.bioz.com/result/longamp taq/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    longamp taq - by Bioz Stars, 2021-07
    99/100 stars

    Images

    1) Product Images from "Rapid bacterial identification by direct PCR amplification of 16S rRNA genes using the MinION™ nanopore sequencer"

    Article Title: Rapid bacterial identification by direct PCR amplification of 16S rRNA genes using the MinION™ nanopore sequencer

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12590

    Taxonomic assignment of the mock community consisting of 10 bacterial species. A mixture of DNA from 10 different bacterial species was analyzed by 16S rRNA amplicon sequencing using MinION™. PCR amplification was performed with LongAmp™ Taq or KAPA2G™ polymerase. The samples were analyzed by MinION™ sequencing with different run time conditions, and the percentage of reads mapping to the 10 bacterial species is shown. Expected abundance of individual taxa is based on the genome size and copy number of 16S rRNA genes.
    Figure Legend Snippet: Taxonomic assignment of the mock community consisting of 10 bacterial species. A mixture of DNA from 10 different bacterial species was analyzed by 16S rRNA amplicon sequencing using MinION™. PCR amplification was performed with LongAmp™ Taq or KAPA2G™ polymerase. The samples were analyzed by MinION™ sequencing with different run time conditions, and the percentage of reads mapping to the 10 bacterial species is shown. Expected abundance of individual taxa is based on the genome size and copy number of 16S rRNA genes.

    Techniques Used: Amplification, Sequencing, Polymerase Chain Reaction

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Short and Long-Read Sequencing Survey of the Dynamic Transcriptomes of African Swine Fever Virus and the Host Cells
    Article Snippet: The strand-switching buffer mixture (RT buffer, RNaseOUT, nuclease-free water, and SSP) was added to the samples, which were incubated at 40°C for 2 min. RT was conducted by adding Maxima H Minus Reverse Transcriptase at 42°C for 90 min. .. The enzyme was inactivated by increasing the temperature to 85°C for 5 min. LongAmp Taq Master Mix, one of the Low Input barcode primers (LWB01-12, from the ONT's SQK-PBK004 kit, ), and nuclease-free water were included in the RT reaction mixture. shows the PCR conditions. .. PCR products were treated with exonuclease (NEB), and the mixture was then incubated at 37°C for 15 min, followed by 80°C for 15 min. AMPure Beads (0.8 BR) was used for purification, and the clean sample was eluted.

    Article Title: Rapid bacterial identification by direct PCR amplification of 16S rRNA genes using the MinION™ nanopore sequencer
    Article Snippet: Bacterial DNA was purified using DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) and used as a PCR template. .. PCR amplification of 16S rRNA genes was conducted using the 16S Barcoding Kit (SQK‐RAB204; Oxford Nanopore Technologies, Oxford, UK) containing the 27F/1492R primer set , and LongAmp™ Taq 2× Master Mix (New England Biolabs, Ipswich, MA, USA). .. Amplification was performed using an Applied Biosystems Veriti™ Thermal Cycler (Thermo Fischer Scientific, Waltham, MA, USA) with the following PCR conditions: initial denaturation at 95 °C for 3 min, 25 cycles of 95 °C for 20 s, 55 °C for 30 s, and 65 °C for 2 min, followed by a final extension at 65 °C for 5 min. To determine the effects of human DNA contamination on 16S rRNA gene amplification, genomic DNA purified from the human monocytic cell line THP‐1 was mixed with E. coli DNA and subjected to PCR.

    Article Title: Microtubule nucleation promoters Mto1 and Mto2 regulate cytokinesis in fission yeast
    Article Snippet: Lithium acetate transformation was used to introduce 5–10 μg of integration cassette DNA ( ). .. Integration at the correct locus was verified by colony PCR using LongAmp Taq 2X Master Mix (New England BioLabs), and the mto2 gene was sequenced in its entirety to verify the S338N or S338C point mutations. .. The bgs1D277N strain was generated by CRISPR ( ).

    Article Title: Randomly primed, strand-switching MinION-based sequencing for the detection and characterization of cultured RNA viruses
    Article Snippet: The cDNA was purified by KAPA Pure Beads (Kapa Biosystems, Wilmington, MA) or with AMPure XP beads (Beckman Coulter, Indianapolis, IN) at 0.7× beads: solution ratio. .. Barcoding PCRThe reverse-transcribed cDNA was amplified following ONT’s 1D PCR barcoding cDNA (SQK-LSK108) protocol using the PCR Barcoding Expansion 1-12 (EXP-PBC001) kit (ONT) and LongAmp Taq 2× Master Mix (New England Biolabs, MA) with the following thermocycling conditions: 95°C for 3 minutes; 18 cycles of 95°C for 15 seconds, 62°C for 15 seconds, 65°C for 22 minutes; 65°C for 23 minutes. ..

    Article Title: Enrichment by hybridisation of long DNA fragments for Nanopore sequencing
    Article Snippet: .. The ligated DNA was amplified using Long Amp Taq 2x Master mix (#M0287, NEB) and ONT PCR primers with the following program: 95 °C 3 min; 15–18 cycles of 95 °C 15 sec, 62 °C 15 sec, 65 °C 10 min; 65 °C 20 min; 4 °C hold. .. A second round of end repair and dA-tailing was performed on 500 ng of enriched, amplified PCR product using SureSelectXT reagents as described above, but without purification after dA-tailing.

    Article Title: Short and long-read genome sequencing methodologies for somatic variant detection; genomic analysis of a patient with diffuse large B-cell lymphoma
    Article Snippet: Subsequently, full-length double stranded cDNA was generated employing the Nanopore Strand Switching Primer and amplified in 50 µl reaction volumes. .. The PCR reaction contained 5 µl cDNA, 25 µl 2X LongAmp Taq Master Mix (NEB Biolabs Inc, Ipswich, Massachusetts, USA, cat. #M0287S), 1.5 µl Nanopore cDNA Primers and 18.5 µl NFW. .. The amplification products were normalised to ~ 400 fmol into 20 µl of Nanopore Rapid Annealing Buffer.

    Article Title: Targeted RNA-Based Oxford Nanopore Sequencing for Typing 12 Classical HLA Genes
    Article Snippet: Using this information, the molarity of each pool was calculated using the DNA concentration (ng/μl) and the average fragment length (bp) of the gene pool. .. ONT Library Preparation and Sequencing ONT’s sequencing compatible barcoded fragments were prepared in a PCR reaction 0.5 nM of DNA from gene pools, 2 μl of PCR barcode from the 96 PCR Barcoding Kit (ONT), 50 μl of LongAmp Taq 2x Mix (New England Biolabs), and Nuclease-Free water in a final volume of 100 μl where ONT’s universal tails were used as a template for barcode introducing primers. .. The PCR was performed in the following conditions: initial denaturation of 3 min at 95°C, following 15 cycles of 15 s at 95°C, 15 s at 62°C, 30 s at 65°C, and a final extension step 3 min at 65°C.

    Amplification:

    Article Title: Rapid bacterial identification by direct PCR amplification of 16S rRNA genes using the MinION™ nanopore sequencer
    Article Snippet: Bacterial DNA was purified using DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) and used as a PCR template. .. PCR amplification of 16S rRNA genes was conducted using the 16S Barcoding Kit (SQK‐RAB204; Oxford Nanopore Technologies, Oxford, UK) containing the 27F/1492R primer set , and LongAmp™ Taq 2× Master Mix (New England Biolabs, Ipswich, MA, USA). .. Amplification was performed using an Applied Biosystems Veriti™ Thermal Cycler (Thermo Fischer Scientific, Waltham, MA, USA) with the following PCR conditions: initial denaturation at 95 °C for 3 min, 25 cycles of 95 °C for 20 s, 55 °C for 30 s, and 65 °C for 2 min, followed by a final extension at 65 °C for 5 min. To determine the effects of human DNA contamination on 16S rRNA gene amplification, genomic DNA purified from the human monocytic cell line THP‐1 was mixed with E. coli DNA and subjected to PCR.

    Article Title: Randomly primed, strand-switching MinION-based sequencing for the detection and characterization of cultured RNA viruses
    Article Snippet: The cDNA was purified by KAPA Pure Beads (Kapa Biosystems, Wilmington, MA) or with AMPure XP beads (Beckman Coulter, Indianapolis, IN) at 0.7× beads: solution ratio. .. Barcoding PCRThe reverse-transcribed cDNA was amplified following ONT’s 1D PCR barcoding cDNA (SQK-LSK108) protocol using the PCR Barcoding Expansion 1-12 (EXP-PBC001) kit (ONT) and LongAmp Taq 2× Master Mix (New England Biolabs, MA) with the following thermocycling conditions: 95°C for 3 minutes; 18 cycles of 95°C for 15 seconds, 62°C for 15 seconds, 65°C for 22 minutes; 65°C for 23 minutes. ..

    Article Title: Enrichment by hybridisation of long DNA fragments for Nanopore sequencing
    Article Snippet: .. The ligated DNA was amplified using Long Amp Taq 2x Master mix (#M0287, NEB) and ONT PCR primers with the following program: 95 °C 3 min; 15–18 cycles of 95 °C 15 sec, 62 °C 15 sec, 65 °C 10 min; 65 °C 20 min; 4 °C hold. .. A second round of end repair and dA-tailing was performed on 500 ng of enriched, amplified PCR product using SureSelectXT reagents as described above, but without purification after dA-tailing.

    Sequencing:

    Article Title: Targeted RNA-Based Oxford Nanopore Sequencing for Typing 12 Classical HLA Genes
    Article Snippet: Using this information, the molarity of each pool was calculated using the DNA concentration (ng/μl) and the average fragment length (bp) of the gene pool. .. ONT Library Preparation and Sequencing ONT’s sequencing compatible barcoded fragments were prepared in a PCR reaction 0.5 nM of DNA from gene pools, 2 μl of PCR barcode from the 96 PCR Barcoding Kit (ONT), 50 μl of LongAmp Taq 2x Mix (New England Biolabs), and Nuclease-Free water in a final volume of 100 μl where ONT’s universal tails were used as a template for barcode introducing primers. .. The PCR was performed in the following conditions: initial denaturation of 3 min at 95°C, following 15 cycles of 15 s at 95°C, 15 s at 62°C, 30 s at 65°C, and a final extension step 3 min at 65°C.

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  • 99
    New England Biolabs crimson longamp taq polymerase
    Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB <t>LongAmp</t> <t>Taq</t> polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.
    Crimson Longamp Taq Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/crimson longamp taq polymerase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    crimson longamp taq polymerase - by Bioz Stars, 2021-07
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    Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Journal: Journal of Insect Science

    Article Title: Rapid Identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) Using Ribosomal RNA Internal Transcribed Spacer 1

    doi: 10.1093/jisesa/iev137

    Figure Lengend Snippet: Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Article Snippet: Initial tests of species-specific fragment amplification were carried out using Crimson LongAmp Taq polymerase with a 45 s initial denaturation step at 95°C followed by 35 cycles of 15 s denaturation at 95°C, 10 s annealing at 60°C, and 30 s extension at 72°C.

    Techniques: Produced, Modification, Amplification

    Effect of DNase I treatment on Taq DNA polymerase-mediated RT-qPCR assay. Taq DNA polymerase purchased from NEB was used to operate CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays using SARS-CoV-2 viral genomic RNA (panels A-C) or N gene armored RNA (panels D-F) treated with DNase I. Amplification curves shown in panels A-C resulted from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of SARS-CoV-2 genomic RNA. Amplification curves in panels D-F resulted from 30,000 (black traces), 3,000 (red traces), 300 (blue traces), 30 (pink traces) and 0 (gray traces) copies of N gene armored RNA. Representative Ct values for RT-qPCR amplification of indicated copies of untreated and DNase I treated SARS-CoV-2 genomic RNA and N gene armored RNA are tabulated.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: Effect of DNase I treatment on Taq DNA polymerase-mediated RT-qPCR assay. Taq DNA polymerase purchased from NEB was used to operate CDC SARS-CoV-2 N1, N2, and N3 TaqMan RT-qPCR assays using SARS-CoV-2 viral genomic RNA (panels A-C) or N gene armored RNA (panels D-F) treated with DNase I. Amplification curves shown in panels A-C resulted from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of SARS-CoV-2 genomic RNA. Amplification curves in panels D-F resulted from 30,000 (black traces), 3,000 (red traces), 300 (blue traces), 30 (pink traces) and 0 (gray traces) copies of N gene armored RNA. Representative Ct values for RT-qPCR amplification of indicated copies of untreated and DNase I treated SARS-CoV-2 genomic RNA and N gene armored RNA are tabulated.

    Article Snippet: These results demonstrate that Taq polymerase can generate amplicons from RNA during a normal thermal cycling reaction, and that a pre-incubation step greatly improves detection.

    Techniques: Quantitative RT-PCR, Amplification

    SARS-CoV-2 N1 TaqMan RT-qPCR assays performed using NEB Taq DNA polymerase and N gene armored RNA in indicated buffers. Buffer compositions are detailed in Table 2 . Amplification curves resulting from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces), 30 (green traces), and 0 (gray) copies of SARS-CoV-2 N gene armored RNA are depicted.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: SARS-CoV-2 N1 TaqMan RT-qPCR assays performed using NEB Taq DNA polymerase and N gene armored RNA in indicated buffers. Buffer compositions are detailed in Table 2 . Amplification curves resulting from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces), 30 (green traces), and 0 (gray) copies of SARS-CoV-2 N gene armored RNA are depicted.

    Article Snippet: These results demonstrate that Taq polymerase can generate amplicons from RNA during a normal thermal cycling reaction, and that a pre-incubation step greatly improves detection.

    Techniques: Quantitative RT-PCR, Amplification

    TaqMan RT-qPCR analysis of SARS-CoV-2 viral genomic RNA and RNaseP armored RNA using Taq DNA polymerase-based one-enzyme assays. CDC SARS-CoV-2 N gene assays, N1, N2, and N3, and RNaseP assay were performed using Taq DNA polymerase from either NEB (panels A-H) or Thermo Fisher (panels I-P). Assays were performed either using the companion commercial buffer (panels A-D and panels I-L) or using Gen 6 A buffer (panels E-H and panels M-P). Amplification curves from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of viral genomic RNA are depicted in panels A-C, E-G, I-K, and M-O. Amplification curves from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces) and 0 (gray traces) copies of armored RNaseP RNA are depicted in panes D, H, L, and P.

    Journal: bioRxiv

    Article Title: One enzyme reverse transcription qPCR using Taq DNA polymerase

    doi: 10.1101/2020.05.27.120238

    Figure Lengend Snippet: TaqMan RT-qPCR analysis of SARS-CoV-2 viral genomic RNA and RNaseP armored RNA using Taq DNA polymerase-based one-enzyme assays. CDC SARS-CoV-2 N gene assays, N1, N2, and N3, and RNaseP assay were performed using Taq DNA polymerase from either NEB (panels A-H) or Thermo Fisher (panels I-P). Assays were performed either using the companion commercial buffer (panels A-D and panels I-L) or using Gen 6 A buffer (panels E-H and panels M-P). Amplification curves from 6000 (black traces), 600 (red traces), 60 (blue traces), 6 (pink traces), and 0 (gray traces) copies of viral genomic RNA are depicted in panels A-C, E-G, I-K, and M-O. Amplification curves from 3 × 10 5 (black traces), 3 × 10 4 (red traces), 3 × 10 3 (blue traces), 3 × 10 2 (pink traces) and 0 (gray traces) copies of armored RNaseP RNA are depicted in panes D, H, L, and P.

    Article Snippet: These results demonstrate that Taq polymerase can generate amplicons from RNA during a normal thermal cycling reaction, and that a pre-incubation step greatly improves detection.

    Techniques: Quantitative RT-PCR, Amplification

    DMPK CTG repeat contraction and loss of methylation in the upstream CpG rich region in MSH2KD clonal lines. The top of the graph represents the DMPK CTG repeat region, with the 25 upstream CpG sites (vertical bars) and the CTCF1 site. The methylation levels are shown on the left and CTG repeat length distributions are shown on the right per clonal line of VUB03-DM1 and VUB19-DM1 from passage 4 to passage 20. The X-axis shows the passage number and median CTG repeat size. Methylation is shown for 25 CpG sites and a total of 100 epi-alleles were analyzed per cell passage. Bubble size is relative to the number of epi-alleles with the indicated percentage of CpG methylation, as previously described 19 . The horizontal grey line represents the threshold for non-methylated alleles and is established on methylation levels in non-DM1 individuals as described in Barbé et al. (2017) 19 . All alleles above the threshold are considered as methylated alleles. In the right panel, each dot represents the median (most frequently present) CTG repeat size of one low input LongAmp PCR reaction, spanning the repeat. The horizontal line represents the median for that clonal cell line and passage number. Abbreviations: pas.: passage

    Journal: bioRxiv

    Article Title: MSH2 knock-down shows CTG repeat stability and concomitant upstream demethylation at the DMPK locus in myotonic dystrophy type 1 human embryonic stem cells

    doi: 10.1101/2020.09.25.313197

    Figure Lengend Snippet: DMPK CTG repeat contraction and loss of methylation in the upstream CpG rich region in MSH2KD clonal lines. The top of the graph represents the DMPK CTG repeat region, with the 25 upstream CpG sites (vertical bars) and the CTCF1 site. The methylation levels are shown on the left and CTG repeat length distributions are shown on the right per clonal line of VUB03-DM1 and VUB19-DM1 from passage 4 to passage 20. The X-axis shows the passage number and median CTG repeat size. Methylation is shown for 25 CpG sites and a total of 100 epi-alleles were analyzed per cell passage. Bubble size is relative to the number of epi-alleles with the indicated percentage of CpG methylation, as previously described 19 . The horizontal grey line represents the threshold for non-methylated alleles and is established on methylation levels in non-DM1 individuals as described in Barbé et al. (2017) 19 . All alleles above the threshold are considered as methylated alleles. In the right panel, each dot represents the median (most frequently present) CTG repeat size of one low input LongAmp PCR reaction, spanning the repeat. The horizontal line represents the median for that clonal cell line and passage number. Abbreviations: pas.: passage

    Article Snippet: Twenty to 50pg of DNA was amplified in a 25 µL reaction mix containing 2.5 units LongAmp Taq DNA polymerase, 1x LongAmp buffer (New England Biolabs), 0.2 mM dNTPs (Illustra DNA polymerization mix, GE Healthcare) and 0.4 µM of primers DM101 and DM102 (Integrated DNA Technologies) and 2.5% dimethyl sulphoxide (DMSO).

    Techniques: Methylation, CpG Methylation Assay, Polymerase Chain Reaction

    Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Article Snippet: LongAmp Taq DNA polymerase (New England Biolabs) was used in our experiments with great success.

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control

    Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Article Snippet: LongAmp Taq DNA polymerase (New England Biolabs) was used in our experiments with great success.

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control