colorimetric warmstart lamp 2x master mix  (New England Biolabs)

Journal: Microorganisms

Article Title: Development and Validation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Glaesserella (Haemophilus) parasuis

doi: 10.3390/microorganisms9010041

Figure Lengend Snippet: Results of the WarmStart Colorimetric LAMP 2X Master Mix assay for detection of LAMP amplicons with the naked eye. Samples 1–8: serial dilutions of DNA from strain G. ( H.) parasuis DSM 21448 starting at concentrations of 10 ng/µL up to 1 fg/µL. Sample 9: DNA from Actinobacillus minor CCUG 38923 T . Sample 10: no template control.

Article Snippet: The isothermal OptiGene Isothermal Master Mix was replaced by the colorimetric WarmStart LAMP 2X Master Mix (New England BioLabs, Frankfurt am Main, Germany).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Colorimetric RT-LAMP for COVID-19 diagnosis validation using 100 clinical samples. Clinical samples were collected from symptomatic and hospitalized patients by nasopharyngeal swabs in partnership with CT-Vacinas/UFMG, Belo Horizonte, Brazil. Samples were obtained from different parts including Brazilian Southeast and Northeast regions. The reaction was performed at 65°C during 30 min using WarmStart ® colorimetric LAMP master mix (NEB #M1800) in 20 μL final volume. The RT-LAMP reaction targeted SARS-CoV-2 N gene. Yellow content indicates positive reaction, whereas the pink pattern reveals nonreagent samples. Amplicons were resolved in 2% agarose gel and stained with GelRed ® (Biotium #41003) to confirm DNA amplification. Latter pattern confirmed specific SARS-CoV-2 amplification that matches with yellow output tubes, which is not observed in pink nonreagent tests. +C, positive control using RNA extracted from laboratory-Vero E6 cultured inactivated SARS-CoV-2; NTC, nontemplate control. Clinimetric parameters from these samples are presented in Supplementary Figure S1 .

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: Agarose Gel Electrophoresis, Staining, Amplification, Positive Control, Cell Culture

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Colorimetric RT-LAMP allows the detection of SARS-CoV-2 VOCs and VOIs. RT-LAMP reaction was performed at 65°C for 30 min, using the WarmStart ® colorimetric LAMP 2× master mix (NEB #M1804), using multiplex N 2/ E 1 primer sets. The amplicons were migrated in agarose gel at 2% to confirm amplification, as indicated by the characteristic ladder highlighted by GelRed ® staining. NTC, nontemplate control; CS, clinical sample; and +C, positive control. The top panel shows a schematic representation of SARS-CoV-2 spike protein (upper) and where the main mutations are highlighted and represented in SARS-CoV-2 virions (right hand side) present in VOC gamma (B.1), delta (B.1.167.2), and VOI zeta (P.2). The VOCs alpha (B.1.1.7) and beta (B.1.3.51), first reported in the United Kingdom and South Africa, respectively, are also represented. K417N: lysine-to-asparagine substitution at position 417 of spike protein at the receptor biding domain (RBD); V445A: valine-to-alanine substitution at position 445 and so on. L, leucine; Q, glutamine; E, glutamic acid; Y, tyrosine; T, threonine; P, proline; H, histidine; D, aspartic acid; S, serine; F, phenylalanine. del, deletion. Segments of SARS-CoV-2 protein NTD, N-terminal domain; CTD2, C-terminal domain 2 or C terminus of S1 fragment after furin cleavage; FP, fusion peptide; HR1, heptad repeat region 1. SARS-CoV-2 variants were previously sequenced. Variants of interest B.1.1.371 and B.1.1.374 were first reported in Saudi Arabia and Finland, respectively, ( https://cov-lineages.org/ ). Created with biorender.com .

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: Multiplex Assay, Agarose Gel Electrophoresis, Amplification, Staining, Positive Control

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Analytical sensitivity as revealed by the limit of detection (LoD). RNA was extracted from VTM-nasopharyngeal swab, and the genome viral copies input was calculated based on SARS-CoV-2 E gene-harboring plasmid (Bioclin #K228-1) calibration curve. RT-LAMP reaction was performed at 65°C during 30 min using WarmStart ® colorimetric master LAMP mix (NEB #M1800) in 20 μL final volume (upper panel). Amplicons were resolved in 2% agarose gel and stained with GelRed ® (Biotium #41003) to confirm DNA amplification (bottom panel). cps/μL, viral genome copies per microliter; NTC, nontemplate control; VTM, viral transport medium (Bioclin #G092-1).

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: Plasmid Preparation, Agarose Gel Electrophoresis, Staining, Amplification

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Microbial cross-reactivity assay to test SARS-CoV-2 RT-LAMP analytical sensitivity. The test was performed using potentially cross-reacting respiratory viruses or local occurring arboviruses. RT-LAMP reaction was performed at 65°C during 30 min, with additional 10 min, to confirm the absence of cross-reactivity when targeting SARS-CoV-2 E and N genes. The assay was performed using the WarmStart ® colorimetric LAMP 2× master mix (NEB #M1800). Yellow (positive) reaction is observed only when the template is SARS-CoV-2 viral RNA. hRSV, human respiratory syncytial virus; NTC, nontemplate control; M, molecular size marker. RT-LAMP amplification products were resolved in 2% agarose gel and stained with GelRed ® (Biotium #41003) to confirm DNA amplification. DENV3, dengue virus serotype 3; ZIKV, Zika virus; CHIKV, Chikungunya virus; YFV, yellow fever virus; Influenza A (H1N1/H3N2); and influenza B (Yamagata/Victoria).

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: Marker, Amplification, Agarose Gel Electrophoresis, Staining

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Colorimetric RT-LAMP for SARS-CoV-2 detection using genes N , E , and RdRp as target. Selected SARS-CoV-2–positive clinical samples by RT-qPCR were classified as low (Ct 18.9 and 21.7), medium (Ct 26.6 and 28.4), and high (Ct 31.6 and 35.2) Ct values for E gene. They were included as input for colorimetric RT-LAMP reaction using primers targeting N , RdRp (A) , and E genes (B) . RT-LAMP SARS-CoV-2 false-negative samples were more frequent when using E and RdRp genes as target (C) . RT-LAMP reaction was performed at 65°C during 30 min, using the WarmStart ® colorimetric LAMP 2× master mix (NEB #M1800). RT-LAMP amplification products were resolved in 2% agarose gel and stained with GelRed ® (Biotium #41003) to confirm DNA amplification. +C, positive control using SARS-CoV-2 RNA extracted from laboratory-cultured inactivated SARS-CoV-2; NTC, nontemplate control.

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: Quantitative RT-PCR, Amplification, Agarose Gel Electrophoresis, Staining, Positive Control, Cell Culture

Journal: Frontiers in Microbiology

Article Title: Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern

doi: 10.3389/fmicb.2021.713713

Figure Lengend Snippet: Colorimetric RT-LAMP to detect SAR-CoV-2 in RNA extraction–free clinical samples (A) or laboratory-cultured virus (B) . Clinical samples were derived from nasopharyngeal swabs placed on guanidine-containing viral transport medium, diluted 1:10. The RT-PCR Ct values for SARS-CoV-2 based on E gene are as follows: CS134 = 31.8, CS135 = 15.3, CS138 = 18.4, CS139 = 21.7, and CS140 = 24.6. RT-LAMP reaction was performed in 20 μL final volume, incubated at 65°C during 30, 40, or 50 min (inactivated virus) using WarmStart ® colorimetric LAMP master mix (NEB #M1800). Both clinical samples and viruses are RNA extraction–free samples. The amplification products (amplicons) were migrated in agarose gel at 2% to confirm amplification, as indicated by the characteristic ladder highlighted by GelRed ® staining. NTC, nontemplate control; CS, clinical sample; and +C, positive control.

Article Snippet: RT-LAMP reactions were performed according to NEB recommendations, containing the following components: 10 μL of WarmStart® Colorimetric LAMP 2× Master Mix [NEB #M1800 or #M1804, the latter contains dUTP UDG (uracil-DNA-glycosylase) to avoid carryover contamination; composition of both are NEB’s proprietary]—ready-to-use mixture of WarmStart® Bst 2.0 DNA polymerase and WarmStart® RTx (reverse transcriptase for one-step transcription/amplification reaction) in presence of a pH sensor that turns from fuchsia (pink) to yellow in presence of increased proton (acid pH) during DNA polymerization on isothermal amplification, 1.6 μmol/L forward inner/backward inner primers (FIP/BIP); 0.2 μmol/L forward and backward outer primers (F3/B3), and 0.4 μmol/L loop forward and loop backward primers (LF/LB); Ultra-pureTM DNAse/RNase-free distilled water (InvitrogenTM #10977015) was added in quantity enough to complete the final volume reaction of 20 μL; isothermal amplification was performed on VeritiTM thermal cycler (Applied Biosystems, Foster City, CA, United States) at 65°C for 30 min. From clinical samples in the first batch, we used as input, 1 μL of RNA extracted from nasopharyngeal swab placed on guanidine-containing VTM, whereas upon optimization, 5 μL source template was considered from the samples in the second group.

Techniques: RNA Extraction, Cell Culture, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Incubation, Amplification, Agarose Gel Electrophoresis, Staining, Positive Control

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: Detection sensitivity of FAW gBlock dsDNA amplicons (upper), evaluating amount of FAW DNA with gBlock DNA (lower). ( a ) Amplification profile with gBlock templates ranging from 10 8 to 10 copies at ten-fold dilution. ( b ) Anneal derivative of gBlock LAMP amplicons, with an anneal derivative of 81 °C. ( c ) Amplification profile of four-fold dilution of FAW DNA (VAITC 10726) and gBlock DNA (10 5 copies, pink). ( d ) Anneal derivative of LAMP amplicons showing two peaks, 78.5 °C for FAW DNA dilutions and 81 °C for gBlock DNA (pink).

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Amplification

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: Mitochondrial COI DNA sequence (3′ region) alignment showing FAW LAMP primers. Sequence of FAW (grey shading, from Kim et al . 37 ) and other closely related Spodoptera species 20 , 55 obtained from GenBank. Reverse primers are underlined; FIP (5′-3′) is made by combining F1 (reverse compliment) and F2; BIP (5′-3′) is made by combining B1 and B2 (reverse compliment).

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Sequencing

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: Optimised LAMP assay performed on FAW larva and adult moth laboratory DNA extracts. ( a ) Amplification profile, with 7 positive samples amplifying in approx. 10 min and negative sample (dark blue) showing a flat line. ( b ) Anneal derivative of LAMP amplicons, with an anneal derivative of 78.5 °C.

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Lamp Assay, Amplification

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: Time-series of FAW LAMP (new assay primers) using colorimetric master mix. Ninety minutes total amplification time shown in increments of 15 min. Samples: (1) Spodoptera frugiperda , (2) Spodoptera litura (PNG), (3) Spodoptera exigua, (4) Helicoverpa armigera conferta, (5) Mythimna convecta, (6) Leucania loreyi, (7) no-template negative control and (8) FAW gBlock DNA dilution 10 6 . The colour change from pink to yellow in tube 1 and 8 indicates positive samples. Negative samples did not change colour.

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Amplification, Negative Control

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: Maximum Likelihood tree (5′-COI DNA sequences) of samples used for testing FAW LAMP assay. Bootstrap values indicated on nodes. AgVic, Agricultural Victoria; CSIRO, Commonwealth Scientific and Industrial Research Organisation; QDAF, Department of Agriculture and Fisheries Queensland; Vic, Victoria Australia; Qld, Queensland Australia; PNG, Papua New Guinea.

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Lamp Assay

Journal: Scientific Reports

Article Title: A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

doi: 10.1038/s41598-021-04496-x

Figure Lengend Snippet: DNA sensitivity test of FAW LAMP and FAW real-time PCR assays. ( a,b ) A four-fold DNA dilution series of two biological replicates of FAW larvae (VAITC 10707 and 10726) DNA amount ranging from 40.0 ng/µL to 2.441 × 10 –3 ng/µL. ( a ) FAW LAMP assay amplification time, sensitive to all 8 DNA dilutions tested. ( b ) Real-time PCR Cq values sensitive to all 8 dilutions tested. ( c,d ) A four-fold DNA dilution series of two biological replicates of FAW adult moth (VAITC 10728 and 10729) DNA amount ranging from 1.0 ng/µL to 6.1 × 10 –5 ng/µL. ( c ) FAW LAMP assay amplification time, sensitive to only 5 out of 8 DNA dilutions tested. ( d ) Real-time PCR Cq values sensitive to only 5 out of 8 DNA dilutions tested. Black and white circles represent biological replicate DNA samples.

Article Snippet: We also tested our FAW LAMP assay primers using an alternative colorimetric LAMP master mix (WarmStart Colorimetric LAMP 2 × master mix, DNA & RNA, New England Biolabs Inc.) following published protocols .

Techniques: Real-time Polymerase Chain Reaction, Lamp Assay, Amplification

Journal: Biosensors & bioelectronics

Article Title: Fully 3D Printed Integrated Reactor Array for Point-of-Care Molecular Diagnostics

doi: 10.1016/j.bios.2018.03.009

Figure Lengend Snippet: Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

Article Snippet: Bovine serum albumin (BSA), Poly (ethylene glycol) 8000 (PEG), poly(vinyl alcohol) (PVA) are from Sigma-Aldrich, RT-PCR grade water from Ambion, Inc., intercalating Eva Green fluorescent dye from Biotium, Isothermal Master Mix used in LAMP reaction buffer from ISO-001nd, OptiGene, Horsham, UK, WarmStart® colorimetric LAMP 2X master mix from New England Biolabs Inc. N. meningitidis (ATCC 13098) from American Type Culture Collection (ATCC, Rockville, MD).

Techniques: Fluorescence, Lamp Assay, Chromatin Immunoprecipitation, Amplification, Concentration Assay