Author: Barnard, Karen N.; Wasik, Brian R.; LaClair, Justin R.; Buchholz, David W.; Weichert, Wendy S.; Alford-Lawrence, Brynn K.; Aguilar, Hector C.; Parrish, Colin R.
Title: Expression of 9-O- and 7,9-O-Acetyl Modified Sialic Acid in Cells and Their Effects on Influenza Viruses Document date: 2019_12_3
ID: 11ejfiwe_32
Snippet: qPCR of SIAE and CasD1 expression. RNA from cells was extracted by using EZNA total RNA kit I (Omega Bio-Tek), and cDNA was synthesized using the SuperScript II reverse transcriptase (Invitrogen) standard protocol with oligo(dT) 12-18 primers (Invitrogen). CasD1-and SIAE-specific primers were designed using Geneious (Biomatters, Ltd.). For CasD1, adequate primers could not be targeted around the CRISPR-Cas9 edit site due to the high G/C content. .....
Document: qPCR of SIAE and CasD1 expression. RNA from cells was extracted by using EZNA total RNA kit I (Omega Bio-Tek), and cDNA was synthesized using the SuperScript II reverse transcriptase (Invitrogen) standard protocol with oligo(dT) 12-18 primers (Invitrogen). CasD1-and SIAE-specific primers were designed using Geneious (Biomatters, Ltd.). For CasD1, adequate primers could not be targeted around the CRISPR-Cas9 edit site due to the high G/C content. qPCR was performed on a Applied Biosystems StepOnePlus real-time PCR system using Fast SYBR green (Bio-Rad). Data were analyzed using StepOne software (Applied Biosystems, v2.1) and GraphPad Prism statistical analysis software.
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