Equal amounts of cell cultures, corresponding to 2x105 cells, were collected at the appropriate time points following infection (hours post-infection, hpi) and processed by using the EZ1 viral nucleic acid kit on a EZ1 platform (Qiagen), following the manufacturer’s instructions, in order to obtain a total nucleic acid fraction, containing both viral DNA and RNA in elution volumes of 120 μL. Volumes of 10 μL were then used in the subsequent qPCR and qRT-PCR assays for the quantitative evaluation of target viral nucleic acids. For quantitative analysis of viral DNA, aliquots of the eluted nucleic acids were directly amplified in a qPCR assay. For quantitative analysis of viral RNA, corresponding aliquots were first treated with Turbo DNAfree reagent (Ambion) and then amplified in a qRT-PCR assay.
Ez1 viral nucleic acid kit
Manufactured by Qiagen
The EZ1 viral nucleic acid kit is a laboratory equipment product designed for the automated extraction and purification of viral nucleic acids, including DNA and RNA, from various sample types. The kit utilizes magnetic bead-based technology to efficiently isolate and concentrate the target nucleic acids for downstream analysis.
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Lab products found in correlation
2 protocols using ez1 viral nucleic acid kit
1
Quantitative Viral Nucleic Acid Analysis
2
Viral Nucleic Acid Extraction and qPCR
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Equal amounts of cell cultures, corresponding to 1.5×10 5 cells and to respective cell-free supernatants, collected at the appropriate time points following transfection or infection were processed by using the EZ1 Viral Nucleic Acid Kit on a EZ1 platform (Qiagen), following the manufacturer's instructions, in order to obtain a total nucleic acid fraction in elution volumes of 150 µL. Volumes of 10 µL were then used in the subsequent qPCR assays for the quantitative evaluation of target viral nucleic acids. For the analysis of supernatants, a benzonase treatment was also carried out by incubating 10 µL of supernatants with 1 µL of enzyme (Novagen), for 4 h at 37 °C, before nucleic acid purification.
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