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Comparison of protocols for removal of melanin from genomic DNA to optimize PCR amplification of DNA purified from highly pigmented lesions.

Melanin is produced by melanocytes and protects against DNA damage by ultraviolet light. Unfortunately, the melanin protein present in melanoma tumor cells is often co-purified during DNA extraction, and this contamination may inhibit subsequent PCR methods, which directly impacts research applications and the molecular diagnostic tests needed for targeted therapeutics. There are presently no described purification protocols that efficiently remove melanin from genomic DNA. In this study, we compare six different methods for melanin removal from genomic DNA: Agarose Gel Electrophoresis, 1mg Chelex®-100, Chelex®-100 5%, centrifugation, OneStep™ PCR Inhibitor Removal Kit and centrifugation plus OneStep™ PCR Inhibitor Removal Kit. Each comparison was made using 16 formalin-fixed paraffin-embedded (FFPE) and 11 fresh cell line samples. All samples were initially tested using the multiplex PCR reaction for GAPDH gene that generates different sized amplified products: 100, 200, 300 and 400 base pairs, which could be inhibited by the addition of exogenous melanin. Six purification protocols were then applied, and all samples that amplified at least one GAPDH fragment were sequenced to analyze the presence of the BRAF V600E mutation. The efficiencies of amplification decreased for larger sized fragments in all methods. Our comparisons showed that centrifugation combined with the OneStep™ PCR Inhibitor Removal Kit was superior to all other methods for successful BRAF sequencing with 100% (100bp), 75% (200bp), 50% (300bp), and 31.3% (400bp) amplification efficiencies for the different amplicon sizes. In conclusion, this genomic DNA extraction method is highly efficient for successful PCR when tumor samples are contaminated with melanin.

1411 related Products with: Comparison of protocols for removal of melanin from genomic DNA to optimize PCR amplification of DNA purified from highly pigmented lesions.

Ofloxacin CAS Number [824 Human Dnak (HSP70) His ta Protease, DNASE free hea Protease, DNASE free hea Protease, DNASE free hea DNASE I MOUSE ANTI BOVINE ROTAVIR MOUSE ANTI BORRELIA BURGD DNAI2 antibody Source Rab DNAJB2 antibody Source Ra DNAJC7 antibody Source Ra DNAJA2 antibody Source Ra

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Comparing DNA quantity and quality using saliva collection following food and beverage consumption.

With the democratization of genetic testing, researchers, clinicians, and educators must consider the varying degree of field conditions when collecting samples for genetic analyses. For genotyping or sequencing studies, study designers have multiple options from which to choose, including cheek swabs and saliva sampling. One significant benefit of saliva collection is that it can be done remotely, in the privacy of one's home. This same benefit adds a risk of compliance. Therefore, our goal with this study was to see if the quality and quantity of the saliva collection by a saliva DNA collection kit would be affected by not following the manufacturer's directions, i.e., drinking or eating right before collection.

2969 related Products with: Comparing DNA quantity and quality using saliva collection following food and beverage consumption.

Androgen Receptor (Phosph Androgen Receptor (Phosph Rabbit Anti-Human Androge Rabbit Anti-Human Androge Androgen Receptor (Ab 650 AZD-3514 Mechanisms: Andr 17β-Acetoxy-2α-bromo-5 (5α,16β)-N-Acetyl-16-[2 (5α,16β)-N-Acetyl-16-ac 5α-N-Acetyl-2'H-androst- 5α-N-Acetyl-2'H-androst- 3-O-Acetyl 5,14-Androstad

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Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes.

Transgenic zebrafish lines with the expression of a fluorescent reporter under the control of a cell-type specific promoter, enable transcriptome analysis of FACS sorted cell populations. RNA quality and yield are key determinant factors for accurate expression profiling. Limited cell number and FACS induced cellular stress make RNA isolation of sorted zebrafish cells a delicate process. We aimed to optimize a workflow to extract sufficient amounts of high-quality RNA from a limited number of FACS sorted cells from Tg(fli1a:GFP) zebrafish embryos, which can be used for accurate gene expression analysis.

1851 related Products with: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes.

anti CD38 Hematopoietic p anti Transferrin receptor High density non small ce Multiple lung carcinoma ( MarkerGene™ Cellular Se Fontana-Masson Stain Kit Fontana-Masson Stain Kit Anti C Reactive Protein A anti HSV (II) gB IgG1 (mo anti HCMV IE pp65 IgG1 (m anti HCMV gB IgG1 (monocl Epidermal Growth Factor (

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Evaluation and statistical optimization of a method for methylated cell-free fetal DNA extraction from maternal plasma.

Methylated cell-free fetal DNA (cffDNA) in maternal plasma can potentially be used as a biomarker for accurate noninvasive prenatal testing (NIPT) of fetal disorders. Recovery and purification of cffDNA are key steps for downstream applications. In this study, we aimed to developed and evaluated different aspects of an optimized method and compared its efficiency with common methods used for extraction of methylated cffDNA.

2426 related Products with: Evaluation and statistical optimization of a method for methylated cell-free fetal DNA extraction from maternal plasma.

QuantiChrom™ LDH Cytoto Bovine Androstenedione,AS QuantiChrom™ Formaldehy EnzyChrom™ Free Fatty A AccuPrep Viral RNA Extrac AccuRapid™ Cell Free Pr Mammalian Cell Extraction Mammalian Cell Extraction Multiple lung carcinoma ( MarkerGeneTM TAMRA Antibo MarkerGene™ LysoLive™ EpiQuik Whole Cell Extrac

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Effect of the extraction and purification of soil DNA and pooling of PCR amplification products on the description of bacterial and archaeal communities.

This study evaluated the effects of DNA extraction method, DNA purification and pooling of PCR amplification products on the description of bacterial and archaeal diversity.

1530 related Products with: Effect of the extraction and purification of soil DNA and pooling of PCR amplification products on the description of bacterial and archaeal communities.

Ofloxacin CAS Number [824 Androgen Receptor (Phosph Androgen Receptor (Phosph Rabbit Anti-Human Androge Rabbit Anti-Human Androge Androgen Receptor (Ab 650 AZD-3514 Mechanisms: Andr 17β-Acetoxy-2α-bromo-5 (5α,16β)-N-Acetyl-16-[2 (5α,16β)-N-Acetyl-16-ac 5α-N-Acetyl-2'H-androst- 5α-N-Acetyl-2'H-androst-

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Successful extraction and PCR amplification of Giardia DNA from formalin-fixed stool samples.

Extracting genomic DNA of pathogenic agents from formalin-fixed specimens is inherently difficult. Storage of samples in formalin results in nucleic acid cross-linking and DNA fragmentation. In this study, DNA was extracted from 45 Giardia-positive stool samples stored in formalin and subjected to PCR amplification targeting the triose phosphate isomerase (tpi), beta gardin (bg) and glutamate dehydrogenase (gdh) genes. Samples were rehydrated by using a descending alcohol series before DNA extraction using a commercial kit. This was followed by EDTA-mediated inhibition of DNase activity and prolonged treatment with proteinase K to digest contaminating proteins. DNA was amplified at rates of 64.4% (29/45) at the tpi, 40% (18/45) at the bg and 20% (9/45) at the gdh loci as seen on nested PCR. DNA quality was subsequently tested in a genotyping experiment which produced high-quality sequences at the tpi (41.2%; 12/29) bg (50%; 9/18), and gdh (22.2%; 2/9) loci and enabled differentiation of Giardia strains at the subtype level. The modified extraction protocol was effective at removing inhibitors and reversing cross-linking of DNA. However, PCR amplification was limited to short fragments of DNA which resulted in highest success rate on amplification of the shortest (334 bp) gene fragment tested.

1793 related Products with: Successful extraction and PCR amplification of Giardia DNA from formalin-fixed stool samples.

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Development of RapidHIT ID using NGMSElect™ Express chemistry for the processing of reference samples within the UK criminal justice system.

The RapidHIT ID is a new instrument which has been designed specifically for non-technical users in non-laboratory environments, for example police custody suites or border control. The development studies presented here were performed using the AmpFℓSTR NGMSElect™ Express STR kit with single source reference samples. The system has been successfully optimised for extraction, PCR and electrophoresis. Once optimised studies on sensitivity, in terms of DNA input template, sample repeatability and instrument reproducibility have shown that the instrument is capable of producing robust profiles within an 85min, window, using mock reference buccal samples. This work has been carried out within the validation framework provided by the United Kingdom Forensic Science Regulator and is a key stage in the validation of the RapidHIT ID for use in custody suites within the UK criminal justice system.

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Filter paper-based spin column method for cost-efficient DNA or RNA purification.

We describe herein a method of recharging used commercial spin columns or assembling homemade spin columns using filter paper as binding material for cost-effective, low throughput nucleic acid purification. The efficiency of filter paper-based spin columns was evaluated for purification of nucleic acids from various sources. Following protocols of commercial kits, we found filter paper to be a useful binding material for purification of nucleic acids, including plant genomic DNA, plant total RNA, PCR products, and DNA from agarose gels. However, filter paper has a weak binding affinity to plasmid DNA in tested miniprep protocols. Protocols for the use of filter paper recharged spin columns or homemade spin columns for low throughput purification of plant genomic DNA and total RNA with unused commercial kit buffers or less expensive homemade buffers are presented.

1437 related Products with: Filter paper-based spin column method for cost-efficient DNA or RNA purification.

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MBL2 gene polymorphisms in HHV-8 infection in people living with HIV/AIDS.

Host genetic factors such as MBL2 gene polymorphisms cause defects in the polymerization of MBL protein and result in a functional deficiency and/or in low serum levels that can influence susceptibility to various viral infections. The aim of this study was to estimate the frequency of alleles, genotypes and haplotypes related to -550, -221 and exon 1 polymorphisms of the MBL2 gene and investigate their association with HHV-8 in people living with HIV/AIDS (PLWHA), as well as the impacts on CD4 cell count and HIV viral load in HIV/HHV-8 coinfected and HIV monoinfected patients.

2610 related Products with: MBL2 gene polymorphisms in HHV-8 infection in people living with HIV/AIDS.

DNA (cytosine 5) methyltr Human Epstein-Barr Virus Mouse Epstein-Barr Virus Rat TGF-beta-inducible ea Rat TGF-beta-inducible ea AZD-8055 Mechanisms: mTOR RDEA-119 (BAY-869766) Mec MK-8033 Mechanisms: c-MET AZD-8330 Mechanisms: MEK 4-Amino-3.4-dideoxy-2-C-( 3-(2-Aminoethyl)-N-methyl 5-(Aminomethyl)indole C9H

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Direct blood PCR: TaqMan-probe based detection of the venous thromboembolism associated mutations factor V Leiden and prothrombin c.20210G>A without DNA extraction.

Practically, the initial step of genetic analysis is the extraction of DNA from blood or other cells, which is often time consuming and cost-intensive. We aimed at establishing a real-time PCR protocol for the detection of the venous thromboembolism associated mutations factor V Leiden (F5 c.1691G>A; p.R506Q) and prothrombin (F2) c.20210G>A from whole blood, without DNA extraction.

2745 related Products with: Direct blood PCR: TaqMan-probe based detection of the venous thromboembolism associated mutations factor V Leiden and prothrombin c.20210G>A without DNA extraction.

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