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10.1016/j.talanta.2021.122227 http://scihub22266oqcxt.onion/10.1016/j.talanta.2021.122227 33773731!7898971!33773731     free     free     free Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Talanta 2021 ; 228 (ä): 122227 Nephropedia Template TP {{tp|p=33773731|t=2021. Quantitative analysis of RNA by HPLC and evaluation of RT-dPCR for coronavirus RNA quantification |pdf=|usr=}}{{33773731}} gab.com Text Quantitative analysis of RNA by HPLC and evaluation of RT-dPCR for coronavirus RNA quantification JO: Talanta http://www.kidney.de/pget.php?&tw=1&pmid=33773731 #FOAvip Nucleic acid detection and quantification have been known to be important at various fields, from genetically modified organisms and gene expression to virus detection. For DNA molecules, digital PCR has been developed as an absolute quantification method which is not dependent on external calibrators. While when it comes to RNA molecules, reverse transcription (RT) step must be taken before PCR amplification to obtain cDNA. With different kinds of reverse transcriptase (RTase) and RT reaction conditions being used in laboratory assays, the efficiency of RT process differs a lot which led variety in quantification results of RNA molecules. In this study, we developed HPLC method combined with enzymatic digestion of RNA to nucleotides for quantification of RNA without RT process. This method was metrologically traceable to four nuceloside monophosphate (NMP) Certification Reference Materials of National Institute of Metrology, China (NIMC) for insurance of accuracy. The established method was used to evaluate the reverse transcription digital polymerase chain reaction (RT-dPCR) of three target genes of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) RNA, including open reading frame 1ab (ORF1ab), nucleocapsid protein (N) and envelope protein (E) gene. Three available RT kits had been evaluated and disparities were observed for the RT efficiency varied from 9% to 182%. It is thus demonstrated that HPLC combined with enzymatic digestion could be a useful method to quantify RNA molecules and evaluate RT efficiency. It is suggested that RT process should be optimized and identified in RNA quantification assays. Twit Text FOAVip Quantitative analysis of RNA by HPLC and evaluation of RT-dPCR for coronavirus RNA quantification ????Talanta http://www.kidney.de/pget.php?&tw=1&pmid=33773731 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33773731 #FOAvip English Wikipedia <ref>{{Cite pmid|33773731}}</ref> Quantitative analysis of RNA by HPLC and evaluation of RT-dPCR for coronavirus RNA quantification #MMPMID33773731 Niu C; Dong L; Gao Y; Zhang Y; Wang X; Wang J Talanta 2021[Jun]; 228 (ä): 122227 PMID33773731show ga Nucleic acid detection and quantification have been known to be important at various fields, from genetically modified organisms and gene expression to virus detection. For DNA molecules, digital PCR has been developed as an absolute quantification method which is not dependent on external calibrators. While when it comes to RNA molecules, reverse transcription (RT) step must be taken before PCR amplification to obtain cDNA. With different kinds of reverse transcriptase (RTase) and RT reaction conditions being used in laboratory assays, the efficiency of RT process differs a lot which led variety in quantification results of RNA molecules. In this study, we developed HPLC method combined with enzymatic digestion of RNA to nucleotides for quantification of RNA without RT process. This method was metrologically traceable to four nuceloside monophosphate (NMP) Certification Reference Materials of National Institute of Metrology, China (NIMC) for insurance of accuracy. The established method was used to evaluate the reverse transcription digital polymerase chain reaction (RT-dPCR) of three target genes of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) RNA, including open reading frame 1ab (ORF1ab), nucleocapsid protein (N) and envelope protein (E) gene. Three available RT kits had been evaluated and disparities were observed for the RT efficiency varied from 9% to 182%. It is thus demonstrated that HPLC combined with enzymatic digestion could be a useful method to quantify RNA molecules and evaluate RT efficiency. It is suggested that RT process should be optimized and identified in RNA quantification assays. |*Proteolysis[MESH] |Animals[MESH] |Chromatography, High Pressure Liquid/*methods/standards[MESH] |Coronavirus Nucleocapsid Proteins/genetics[MESH] |Crotalinae[MESH] |Middle East Respiratory Syndrome Coronavirus/chemistry/genetics[MESH] |Phosphodiesterase I/*chemistry[MESH] |Purine Nucleotides/standards[MESH] |Pyrimidine Nucleotides/standards[MESH] |RNA/*analysis/chemistry[MESH] |Reference Standards[MESH]Quantitative analysis of RNA by HPLC and evaluation of RT-dPCR for cor(epmc).pdf DeepDyve Pubget Overpricing