High-efficiency transformation of archaea by direct PCR products with its application to directed evolution of a thermostable enzyme

Hyperthermophilic archaea with distinctive biochemical and physiological traits are necessary organisms for elementary analysis of life science and have nice potential for biotechnological purposes. Nonetheless, low transformation effectivity of international DNA molecules impedes developments in genetic modification instruments and industrial purposes. On this research, we utilized extended overlap extension PCR (POE-PCR) to generate multimeric DNA molecules after which remodeled them into two hyperthermophilic archaea, Thermococcus kodakarensis KOD1 and Pyrococcus yayanosii A1.

This research was the primary instance to exhibit the improved transformation efficiencies of POE-PCR merchandise by an element of roughly 100 for T. kodakarensis KOD1 and eight for P. yayanosii A1, respectively, relative to round shuttle plasmids. Moreover, directed evolution of a modestly thermophilic enzyme, Methanothermococcus okinawensis 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), was carried out to acquire extra steady ones attributable to excessive transformation effectivity of T. kodakarensis. T. kodakarensis harbouring probably the most thermostable MoHMGR mutant can develop within the presence of a thermostable antibiotic simvastatin at 85°C and even greater temperatures. This excessive transformation effectivity approach couldn’t solely assist develop extra hyperthermophilic enzyme mutants through directed evolution but additionally simplify genetical modification of archaea, which may very well be novel hosts for industrial biotechnology.

Turmeric, or Curcuma longa, is usually consumed within the South East Asian nations as a medical product and as meals attributable to its therapeutic properties. Nonetheless, with growing demand for turmeric powder, adulterated turmeric powders combined with different low-cost starch powders, comparable to from corn or cassava, are being distributed by meals suppliers for financial profit. Right here, we developed molecular markers utilizing quantitative real-time PCR to determine adulteration in business turmeric powder merchandise. Chloroplast genes, comparable to matK, atpF, and ycf2, had been used to design species-specific primers for C. longa and Zea mays.

 

Screening of economic meat merchandise from grocery store chains for feline derivatives utilizing SP-PCR-RLFP and lab-on-a-chip

Dedication of feline meat in meals merchandise is a vital difficulty for social, well being, financial and spiritual concern. Therefore this paper documented the applying of species particular polymerase chain reaction-restriction fragment size polymorphism (SP-PCR-RFLP) assay focusing on a short-fragments (69 bp) of mitochondrial cytochrome b (cytb) gene to display screen feline meat in business meat merchandise utilizing lab-on-a-chip. The SP-PCR assay proved its specificity theoretically and experimentally whereas testing with totally different frequent animal, aquatic and plant species of DNA. The feline particular (69 bp, 43- and 26-bp) attribute molecular DNA sample was noticed by SP-PCR and RFLP evaluation.
For assay efficiency, it was examined in three several types of business dummy meat merchandise comparable to frankfurters, nuggets and meatballs and digested with AluI-restriction enzyme. The best sensitivity of the assay utilizing lab-on-a-chip was as little as 0.1 pg or 0.01 % (w/w) in business dummy meat merchandise. We’ve got additionally utilized this assay to display screen three necessary business meat merchandise of six totally different model from six grocery store chains situated at three totally different states of Malaysia. Thus complete 378 samples had been examined to validate the specificity, sensitivity, stability of the assay and utilization of it for business meat product screening. Amongst others, the households Opisthorchiidae and Heterophyidae consists of a number of genera inflicting fish-borne zoonoses and distributed additionally in European International locations and which can be included within the ParaFishControl (Superior Instruments and Analysis Methods for Parasite Management in European farmed fish) H2020 EU challenge. As a result of small measurement of the metacercariae, the infective stage for human, these parasites can’t be detected visually in fish and monitoring requires skilled utility of time-consuming strategies.
The goal of this was to develop a speedy and inexpensive molecular technique primarily based on multiplex PCR for simultaneous identification of metacercariae of the most typical European Opisthorchiid and Heterophyid in fish or fish merchandise.

 High-efficiency transformation of archaea by direct PCR products with its application to directed evolution of a thermostable enzyme

High-efficiency transformation of archaea by direct PCR products with its application to directed evolution of a thermostable enzyme

TaqMan probe primarily based multiplex quantitative PCR assay for dedication of bovine, porcine and fish DNA in gelatin admixture, meals merchandise and dietary dietary supplements.

Detection of animal supplies in gelatin-based merchandise is required to handle non secular and cultural considerations, as a result of porcine and bovine gelatins are prohibited in Halal, Kosher and Hindus shopper items. On this paper, multiplex quantitative polymerase chain response (qPCR) assay utilizing TaqMan probe was developed to discriminate bovine, porcine and fish gelatin species in a single assay platform. The assay was particular to cattle, pigs and fish, having been examined towards 14 non-target species. The restrict of detection, beneath gelatin admixed circumstances, was 0.005 ng/µL. Lastly, a pilot survey was undertaken testing 35 Halal branded processed meals and dietary gadgets. Out of 35 samples, solely two had been discovered to be optimistic for porcine species. The authenticity of those two qPCR merchandise was confirmed by DNA sequencing evaluation, which confirmed 99-100% similarity with Sus scrofa (Wild boar) species.
Dried squid merchandise are widespread in China as a snack meals, facet dishes, or refreshments, and the market attraction might be mirrored by the excessive worth that often reaches 497 RMB per kg. Nonetheless, the absence of harmonisation across the definition of squid, in addition to the issues with visible inspection for processed seafood merchandise, make various species substitution for dried squid merchandise a frequent incidence. The goal of the current research was to use a DNA barcoding method for species identification of 48 dried squid merchandise collected from the most important on-line procuring platform in China. Furthermore, we additionally developed a novel SYBR inexperienced real-time PCR assay (simplex and duplex adopted by a melting curve evaluation) particular for Illex argentinus and Todarodes pacificus primarily based on cytochrome C oxidase subunit I (COI) gene.

Human Kinase Library II

HKIN-II 1 set
EUR 450

Human Drug Detoxication II

HDTX-II 1 set
EUR 645

Human Cytokine Primer Library II

HCA-II 1 set
EUR 450

Mouse Cytokine Primer Library II

MCA-II 1 set
EUR 450

Rat Cytokine Primer Library II

RCA-II 1 set
EUR 548

Human Colon Cancer Primer Library II

HCCP-II 1 set
EUR 548

Human DNA Repair Primer Library II

HDRL-II 1 set
EUR 548

Mouse DNA Repair Primer Library II

MDRL-II 1 set
EUR 645

Human Interferon Type II Signaling Primer Library

HIFN-II 1 set
EUR 548

Human Cancer Driver Gene II Primer Library

HCDG-II 1 set
EUR 645

96-Well Plate PCR Products Purification Kit

BS365 5XPlates, 480prep
EUR 250.31
  • Product category: Molecular Biology Kits/DNA - Extraction/PCR Products

96-Well Plate PCR Products Purification Kit

BS3652 2xPlates, 192prep
EUR 155.16
  • Product category: Molecular Biology Kits/DNA - Extraction/PCR Products

EZ-10 Spin Column PCR Products Purification Kit

BS363 50Preps
EUR 76.64
  • Product category: Molecular Biology Kits/DNA - Extraction/PCR Products

EZ-10 Spin Column PCR Products Purification Kit

BS364 100preps
EUR 95.68
  • Product category: Molecular Biology Kits/DNA - Extraction/PCR Products

EZ-10 Spin Column PCR Products Purification Kit

BS664 250preps
EUR 166.58
  • Product category: Molecular Biology Kits/DNA - Extraction/PCR Products

MULTIPLEX KIT PCR MASTITIS PCR kit

PCR-MPX218-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of MULTIPLEX KIT PCR MASTITIS

MULTIPLEX KIT PCR MASTITIS PCR kit

PCR-MPX218-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of MULTIPLEX KIT PCR MASTITIS

MULTIPLEX KIT PCR Babesia & Theileria PCR kit

PCR-MPX401-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of MULTIPLEX KIT PCR Babesia & Theileria

MULTIPLEX KIT PCR Babesia & Theileria PCR kit

PCR-MPX401-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of MULTIPLEX KIT PCR Babesia & Theileria

Halobacillus PCR kit

PCR-A635-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Halobacillus

Halobacillus PCR kit

PCR-A635-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Halobacillus

Enterovirus PCR kit

PCR-H433-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Enterovirus

Enterovirus PCR kit

PCR-H433-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Enterovirus

Mengovirus PCR kit

PCR-H453-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Mengovirus

Mengovirus PCR kit

PCR-H453-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Mengovirus

Astrovirus PCR kit

PCR-H491-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Astrovirus

Astrovirus PCR kit

PCR-H491-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Astrovirus

Calicivirus PCR kit

PCR-H493-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Calicivirus

Calicivirus PCR kit

PCR-H493-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Calicivirus

Adenovirus PCR kit

PCR-H501-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Adenovirus

Adenovirus PCR kit

PCR-H501-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Adenovirus

Polyomavirus PCR kit

PCR-H515-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Polyomavirus

Polyomavirus PCR kit

PCR-H515-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Polyomavirus

Sapovirus PCR kit

PCR-H630-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Sapovirus

Sapovirus PCR kit

PCR-H630-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Sapovirus

Parechovirus PCR kit

PCR-H642-48R 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Parechovirus

Parechovirus PCR kit

PCR-H642-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Parechovirus

Metapneumovirus PCR kit

PCR-H643-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Metapneumovirus

Metapneumovirus PCR kit

PCR-H643-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Metapneumovirus

Entamoeba PCR kit

PCR-H683-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Entamoeba

Entamoeba PCR kit

PCR-H683-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Entamoeba

Acanthamoeba PCR kit

PCR-H684-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Acanthamoeba

Acanthamoeba PCR kit

PCR-H684-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Acanthamoeba

Echovirus PCR kit

PCR-H722-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Echovirus

Echovirus PCR kit

PCR-H722-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Echovirus

ranavirus PCR kit

PCR-V337-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of ranavirus

ranavirus PCR kit

PCR-V337-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of ranavirus

Ruminococcus PCR kit

PCR-V619-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Ruminococcus

Ruminococcus PCR kit

PCR-V619-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Ruminococcus

Bifidobacterium PCR kit

PCR-V621-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Bifidobacterium

Bifidobacterium PCR kit

PCR-V621-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Bifidobacterium

Hantavirus PCR kit

PCR-VH373-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Hantavirus

Hantavirus PCR kit

PCR-VH373-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Hantavirus

Ruminococcus PCR kit

PCR-H619-48D 48T
EUR 453

Ruminococcus PCR kit

PCR-H619-96D 96T
EUR 572

Bifidobacterium PCR kit

PCR-H621-48D 48T
EUR 453

Bifidobacterium PCR kit

PCR-H621-96D 96T
EUR 572

ISOLATE II PCR & Gel Kit

BIO-52058 10 preps Ask for price

ISOLATE II PCR & Gel Kit

BIO-52059 50 preps Ask for price

ISOLATE II PCR & Gel Kit

BIO-52060 250 preps Ask for price

pCR-Blunt II-TOPO-MAPT

PVT17493 2 ug
EUR 231

pCR-Blunt II-TOPO-IFNB1

PVT17536 2 ug
EUR 231

pCR-Blunt II-TOPO-KCNK7

PVT17621 2 ug
EUR 231

48 WELL PCR MICROPLATE

PCR-48-C 10/pk
EUR 102
Description: PCR Plates & Tubes; PCR Strip Tubes and Strip Caps - Axygen

Microthrix parvicella PCR kit

PCR-A575-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Microthrix parvicella

Microthrix parvicella PCR kit

PCR-A575-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Microthrix parvicella

Yersinia pseudotuberculosis PCR kit

PCR-H190-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Yersinia pseudotuberculosis

Yersinia pseudotuberculosis PCR kit

PCR-H190-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Yersinia pseudotuberculosis

Neisseria meningitidis PCR kit

PCR-H421-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Neisseria meningitidis

Neisseria meningitidis PCR kit

PCR-H421-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Neisseria meningitidis

Neisseria sp. PCR kit

PCR-H422-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Neisseria sp.

Neisseria sp. PCR kit

PCR-H422-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Neisseria sp.

Haemophilus influenzae PCR kit

PCR-H423-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Haemophilus influenzae

Haemophilus influenzae PCR kit

PCR-H423-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Haemophilus influenzae

Trypanosoma cruzi PCR kit

PCR-H425-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Trypanosoma cruzi

Trypanosoma cruzi PCR kit

PCR-H425-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Trypanosoma cruzi

Cronobacter sakazakii PCR kit

PCR-H431-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Cronobacter sakazakii

Cronobacter sakazakii PCR kit

PCR-H431-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Cronobacter sakazakii

Clostridium botulinum PCR kit

PCR-H432-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Clostridium botulinum

Clostridium botulinum PCR kit

PCR-H432-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Clostridium botulinum

Virus Zika PCR kit

PCR-H434-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Virus Zika

Virus Zika PCR kit

PCR-H434-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Virus Zika

Corynebacterium diphtheriae PCR kit

PCR-H435-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Corynebacterium diphtheriae

Corynebacterium diphtheriae PCR kit

PCR-H435-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Corynebacterium diphtheriae

Legionella pneumophila PCR kit

PCR-H436-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Legionella pneumophila

Legionella pneumophila PCR kit

PCR-H436-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Legionella pneumophila

Bordetella pertussis PCR kit

PCR-H437-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Bordetella pertussis

Bordetella pertussis PCR kit

PCR-H437-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Bordetella pertussis

Bordetella parapertussis PCR kit

PCR-H438-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Bordetella parapertussis

Bordetella parapertussis PCR kit

PCR-H438-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Bordetella parapertussis

Chlamydia pneumoniae PCR kit

PCR-H439-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Chlamydia pneumoniae

Chlamydia pneumoniae PCR kit

PCR-H439-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Chlamydia pneumoniae

Pneumocystis jiroveci PCR kit

PCR-H440-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Pneumocystis jiroveci

Pneumocystis jiroveci PCR kit

PCR-H440-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Pneumocystis jiroveci

Actinobacillus actinomycetemcomitans PCR kit

PCR-H446-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Actinobacillus actinomycetemcomitans

Actinobacillus actinomycetemcomitans PCR kit

PCR-H446-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Actinobacillus actinomycetemcomitans

Bacteroides fragilis PCR kit

PCR-H447-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Bacteroides fragilis

Bacteroides fragilis PCR kit

PCR-H447-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Bacteroides fragilis

Candida sp. PCR kit

PCR-H448-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Candida sp.

Candida sp. PCR kit

PCR-H448-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Candida sp.

Enterococcus sp. PCR kit

PCR-H449-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Enterococcus sp.

Enterococcus sp. PCR kit

PCR-H449-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Enterococcus sp.

Neisseria gonorrhoeae PCR kit

PCR-H450-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Neisseria gonorrhoeae

Neisseria gonorrhoeae PCR kit

PCR-H450-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Neisseria gonorrhoeae

Dengue virus PCR kit

PCR-H451-48R 50T
EUR 524.4
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Description: An conventional PCR kit for detection of Dengue virus

Dengue virus PCR kit

PCR-H451-96R 100T
EUR 669.4
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Description: An conventional PCR kit for detection of Dengue virus

Chlamydia trachomatis PCR kit

PCR-H452-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Chlamydia trachomatis

Chlamydia trachomatis PCR kit

PCR-H452-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Chlamydia trachomatis

Human papillomavirus PCR kit

PCR-H454-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Human papillomavirus

Human papillomavirus PCR kit

PCR-H454-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Human papillomavirus

Parvovirus В19 PCR kit

PCR-H455-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Parvovirus В19

Parvovirus В19 PCR kit

PCR-H455-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Parvovirus В19

Treponema pallidum PCR kit

PCR-H456-48D 50T
EUR 425.8
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Description: An conventional PCR kit for detection of Treponema pallidum

Treponema pallidum PCR kit

PCR-H456-96D 100T
EUR 521.5
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Description: An conventional PCR kit for detection of Treponema pallidum
Outcomes highlighted the profitable DNA extraction and PCR amplification of a 655 bp COI gene fragment from all merchandise. A most similarity worth within the vary of 98-100% was obtained for all readable sequences utilizing the BOLD and BLAST public databases and 4 species had been recognized.

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