Matches in SemOpenAlex for { <https://semopenalex.org/work/W2006369674> ?p ?o ?g. }
Showing items 1 to 99 of
99
with 100 items per page.
- W2006369674 abstract "With the introduction of the first high-throughput qPCR instrument on the market it became possible to perform thousands of reactions in a single run compared to the previous hundreds. In the high-throughput reaction, only limited volumes of highly concentrated cDNA or DNA samples can be added. This necessity can be solved by pre-amplification, which became a part of the high-throughput experimental workflow. Here, we focused our attention on the limits of the specific target pre-amplification reaction and propose the optimal, general setup for gene expression experiment using BioMark instrument (Fluidigm). For evaluating different pre-amplification factors following conditions were combined: four human blood samples from healthy donors and five transcripts having high to low expression levels; each cDNA sample was pre-amplified at four cycles (15, 18, 21, and 24) and five concentrations (equivalent to 0.078 ng, 0.32 ng, 1.25 ng, 5 ng, and 20 ng of total RNA). Factors identified as critical for a success of cDNA pre-amplification were cycle of pre-amplification, total RNA concentration, and type of gene. The selected pre-amplification reactions were further tested for optimal Cq distribution in a BioMark Array. The following concentrations combined with pre-amplification cycles were optimal for good quality samples: 20 ng of total RNA with 15 cycles of pre-amplification, 20x and 40x diluted; and 5 ng and 20 ng of total RNA with 18 cycles of pre-amplification, both 20x and 40x diluted. We set up upper limits for the bulk gene expression experiment using gene expression Dynamic Array and provided an easy-to-obtain tool for measuring of pre-amplification success. We also showed that variability of the pre-amplification, introduced into the experimental workflow of reverse transcription-qPCR, is lower than variability caused by the reverse transcription step." @default.
- W2006369674 created "2016-06-24" @default.
- W2006369674 creator A5001832574 @default.
- W2006369674 creator A5002417543 @default.
- W2006369674 creator A5013548626 @default.
- W2006369674 creator A5034551341 @default.
- W2006369674 creator A5048335641 @default.
- W2006369674 creator A5081827242 @default.
- W2006369674 creator A5090307974 @default.
- W2006369674 creator A5091649224 @default.
- W2006369674 date "2015-03-11" @default.
- W2006369674 modified "2023-09-26" @default.
- W2006369674 title "Pre-amplification in the context of high-throughput qPCR gene expression experiment" @default.
- W2006369674 cites W165036232 @default.
- W2006369674 cites W1995548146 @default.
- W2006369674 cites W2008594900 @default.
- W2006369674 cites W2023490374 @default.
- W2006369674 cites W2053177037 @default.
- W2006369674 cites W2065848683 @default.
- W2006369674 cites W2074741970 @default.
- W2006369674 cites W2079444664 @default.
- W2006369674 cites W2088925963 @default.
- W2006369674 cites W2093976723 @default.
- W2006369674 cites W2096793378 @default.
- W2006369674 cites W2107726313 @default.
- W2006369674 cites W2131589770 @default.
- W2006369674 cites W2133552723 @default.
- W2006369674 cites W2147241696 @default.
- W2006369674 cites W2148974089 @default.
- W2006369674 cites W2152641162 @default.
- W2006369674 cites W2168420558 @default.
- W2006369674 cites W2169814403 @default.
- W2006369674 cites W2312974961 @default.
- W2006369674 cites W2410897506 @default.
- W2006369674 cites W2502051843 @default.
- W2006369674 cites W4245176872 @default.
- W2006369674 doi "https://doi.org/10.1186/s12867-015-0033-9" @default.
- W2006369674 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4365555" @default.
- W2006369674 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/25888347" @default.
- W2006369674 hasPublicationYear "2015" @default.
- W2006369674 type Work @default.
- W2006369674 sameAs 2006369674 @default.
- W2006369674 citedByCount "33" @default.
- W2006369674 countsByYear W20063696742016 @default.
- W2006369674 countsByYear W20063696742017 @default.
- W2006369674 countsByYear W20063696742018 @default.
- W2006369674 countsByYear W20063696742019 @default.
- W2006369674 countsByYear W20063696742020 @default.
- W2006369674 countsByYear W20063696742021 @default.
- W2006369674 countsByYear W20063696742022 @default.
- W2006369674 countsByYear W20063696742023 @default.
- W2006369674 crossrefType "journal-article" @default.
- W2006369674 hasAuthorship W2006369674A5001832574 @default.
- W2006369674 hasAuthorship W2006369674A5002417543 @default.
- W2006369674 hasAuthorship W2006369674A5013548626 @default.
- W2006369674 hasAuthorship W2006369674A5034551341 @default.
- W2006369674 hasAuthorship W2006369674A5048335641 @default.
- W2006369674 hasAuthorship W2006369674A5081827242 @default.
- W2006369674 hasAuthorship W2006369674A5090307974 @default.
- W2006369674 hasAuthorship W2006369674A5091649224 @default.
- W2006369674 hasBestOaLocation W20063696741 @default.
- W2006369674 hasConcept C104317684 @default.
- W2006369674 hasConcept C150194340 @default.
- W2006369674 hasConcept C153911025 @default.
- W2006369674 hasConcept C187882448 @default.
- W2006369674 hasConcept C48023723 @default.
- W2006369674 hasConcept C54355233 @default.
- W2006369674 hasConcept C67705224 @default.
- W2006369674 hasConcept C86803240 @default.
- W2006369674 hasConceptScore W2006369674C104317684 @default.
- W2006369674 hasConceptScore W2006369674C150194340 @default.
- W2006369674 hasConceptScore W2006369674C153911025 @default.
- W2006369674 hasConceptScore W2006369674C187882448 @default.
- W2006369674 hasConceptScore W2006369674C48023723 @default.
- W2006369674 hasConceptScore W2006369674C54355233 @default.
- W2006369674 hasConceptScore W2006369674C67705224 @default.
- W2006369674 hasConceptScore W2006369674C86803240 @default.
- W2006369674 hasIssue "1" @default.
- W2006369674 hasLocation W20063696741 @default.
- W2006369674 hasLocation W20063696742 @default.
- W2006369674 hasLocation W20063696743 @default.
- W2006369674 hasLocation W20063696744 @default.
- W2006369674 hasOpenAccess W2006369674 @default.
- W2006369674 hasPrimaryLocation W20063696741 @default.
- W2006369674 hasRelatedWork W2009966535 @default.
- W2006369674 hasRelatedWork W2056172754 @default.
- W2006369674 hasRelatedWork W2145879504 @default.
- W2006369674 hasRelatedWork W2365238423 @default.
- W2006369674 hasRelatedWork W2375466314 @default.
- W2006369674 hasRelatedWork W2375593830 @default.
- W2006369674 hasRelatedWork W2380641360 @default.
- W2006369674 hasRelatedWork W2383598698 @default.
- W2006369674 hasRelatedWork W2393389982 @default.
- W2006369674 hasRelatedWork W3103934326 @default.
- W2006369674 hasVolume "16" @default.
- W2006369674 isParatext "false" @default.
- W2006369674 isRetracted "false" @default.
- W2006369674 magId "2006369674" @default.
- W2006369674 workType "article" @default.