A stock of malaria genomic DNA was purified from malaria-positive blood (MPB) using Dynabeads SILANE Viral NA kit (Invitrogen) (Catalog #—37011D) as per the manufacturer’s instructions to be used as an external positive control and standard.
Dynabeads silane viral na kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Dynabeads SILANE Viral NA Kit is a laboratory equipment designed for the isolation and purification of viral nucleic acids (NA) from various sample types. The kit utilizes Dynabeads technology to efficiently capture and extract viral NA, providing a reliable and streamlined process for downstream applications.
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Lab products found in correlation
Quant it picogreen dsdna reagent and kit, by Thermo Fisher Scientific (2 mentions)
Nuclisens lysis buffer, by bioMérieux (1 mentions)
Proteinase k, by Merck Group (1 mentions)
Mineral oil, by Merck Group (1 mentions)
Favorprep tissue genomic dna extraction mini kit, by Favorgen Biotech (1 mentions)
Supersignal west femto maximum sensitivity substrate kit, by Thermo Fisher Scientific (1 mentions)
Maxima h minus cdna synthesis master mix, by Thermo Fisher Scientific (1 mentions)
Favorprep blood cultured cell total rna mini kit, by Favorgen Biotech (1 mentions)
Ficoll density gradient centrifugation kit, by GE Healthcare (1 mentions)
Phosstop inhibitor, by Roche (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Faststart universal sybr green master rox, by Roche (1 mentions)
Salmon sperm dna, by Thermo Fisher Scientific (1 mentions)
6 protocols using dynabeads silane viral na kit
1
Malaria DNA Purification Protocol
2
Microgravity Transition of DNA Extraction Fluids
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We aimed to test how different pertinent test fluids transition between 1 g and low g states in cartridges and wells cut from 96-well DNA extraction plates (VWR 82007-292 Disposable 96 Deep Well Plate, 1.2 mL). The test fluids chosen are as follows: NucliSENS lysis buffer (bioMérieux, Durham, NC, USA) with nucleic acid-binding magnetic particles, NucliSENS wash buffer 1, NucliSENS wash buffer 2, NucliSENS wash buffer 3, NucliSENS elution buffer, Dynabeads lysis buffer (Dynabeads® SILANE Viral NA Kit, Invitrogen, Carlsbad, CA, USA) with nucleic acid-binding magnetic particles, Dynabeads wash buffer 1, and Dynabeads wash buffer 2. The reagents were added to the wells and exposed to brief microgravity during the drops. Wells with significant capillary corner wicking were identified.
3
Optimized RNA Extraction from Samples
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RNA extraction was carried out using the Dynabeads SILANE Viral NA Kit (Thermo Fisher Scientific Inc., MA, USA) according to manufacturer’s instructions with few modifications as following. Briefly, the sample volume was 56 μL (reaction mixture containing only the universal nuclease) and 57.5 μL (reaction mixture containing the enzyme mix), respectively, the wash steps were performed for one time, and the RNA was eluted in a final volume of 60 μL.
4
Circulating Cell-free DNA Extraction
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Extraction of cfDNA from plasma or LNs was performed using Dynabeads SILANE Viral NA kit (Thermo Fisher). For LNs, 50 µL proteinase K (20 mg/mL; Sigma) was added, and LNs were mechanically dissociated using surgical scissors, and then 300 µL lysis/binding buffer was added and mixed prior to cfDNA extraction. The concentration of cfDNA was quantified by Quant‐iT™ PicoGreen® dsDNA reagent and kits (Thermo Fisher).
5
Comprehensive Molecular Analysis Techniques
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Total RNA was extracted using a FavorPrepTM Blood/Cultured Cell Total RNA Mini Kit (Favorgen Biotech, China). Genomic DNA was prepared by using a FavorPrep Tissue Genomic DNA Extraction Mini Kit (Favorgen Biotech, China). For Western blotting, signals were visualized by using a SuperSignal West Femto Maximum Sensitivity Substrate Kit (Thermo, USA). Lymphocyte isolation was performed with a Ficoll density gradient centrifugation kit (GE Healthcare, USA). Cell-free DNA was isolated and quantified by using a Dynabeads® SILANE Viral NA Kit (Thermo, USA) and Quant-iT™ PicoGreen ® dsDNA Reagent and Kits (Thermo, USA).
Reagents and services: LipofectamineTM 3000 (Thermo, USA); TREX1 siRNA (Santa Cruz, USA); cGAS siRNA (Santa Cruz, USA); siRNA control plasmid (Sigma, USA); Mycobacterium tuberculosis (M. tuberculosis; Difco, USA); mineral oil (Sigma, USA); methotrexate (MTX) (LC labs, USA); Cre adeno-associated virus construction (Ubigene, China); TREX1 adeno-associated virus construction (Ubigene, China); dimethyl sulfoxide (DMSO; ACROS, USA); DAPI (Invitrogen, USA); RIPA buffer (10×, Cell Signaling Technology, USA); EDTA-free protease inhibitors, PhosSTOP inhibitor (Roche, Basel, Switzerland); TRIzol (Invitrogen, USA); FastStart Universal SYBR Green Master Rox (Roche Diagnostics, USA); Maxima™ H Minus cDNA Synthesis Master Mix (Thermo, USA); PVDF membranes (Bio-Rad, USA).
Reagents and services: LipofectamineTM 3000 (Thermo, USA); TREX1 siRNA (Santa Cruz, USA); cGAS siRNA (Santa Cruz, USA); siRNA control plasmid (Sigma, USA); Mycobacterium tuberculosis (M. tuberculosis; Difco, USA); mineral oil (Sigma, USA); methotrexate (MTX) (LC labs, USA); Cre adeno-associated virus construction (Ubigene, China); TREX1 adeno-associated virus construction (Ubigene, China); dimethyl sulfoxide (DMSO; ACROS, USA); DAPI (Invitrogen, USA); RIPA buffer (10×, Cell Signaling Technology, USA); EDTA-free protease inhibitors, PhosSTOP inhibitor (Roche, Basel, Switzerland); TRIzol (Invitrogen, USA); FastStart Universal SYBR Green Master Rox (Roche Diagnostics, USA); Maxima™ H Minus cDNA Synthesis Master Mix (Thermo, USA); PVDF membranes (Bio-Rad, USA).
6
Cell-SELEX for RhD Antigen Screening
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The ssDNA sub-libraries in normal saline were further screened by the cell-SELEX strategy described previously, with some modifications (Fig. 1 ) (22 (link),23 (link)). Briefly, the ssDNA sub-libraries were heat-denatured at 95°C for 5 min, snap-cooled and reacted with RhD− RBCs in the presence of 0.1 mg/ml salmon sperm DNA (Thermo Fisher Scientific, Inc.) for 30 min at 25°C. After centrifugation at 1,000 × g for 1 min at 25°C, the supernatants were recovered and incubated with RhD+ RBCs for 30 min at 25°C. The RBCs were magnetized for 2 min using 200 µl MagneLys solution (Diagast) following which they were washed with normal saline five times, and then the ssDNAs bound on the magnetized RBCs were extracted. These were used as templates for PCR amplification in order to generate ssDNA sub-libraries using the Dynabeads SILANE viral NA kit (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. To increase the stringency of selection, the concentrations of ssDNAs and positive selection time were gradually reduced throughout the selection process (Table I ).
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