Journal: Methods and Protocols

Article Title: Comparison of Commercially Available Thermostable DNA Polymerases with Reverse Transcriptase Activity in Coupled Reverse Transcription Polymerase Chain Reaction Assays

doi: 10.3390/mps8010011

Figure Lengend Snippet: Comparison of the indicated enzymes in RT-qPCR with EvaGreen intercalating dye (A) and with TaqMan probe (B). The mean quantification cycles (Cq) ± standard deviation of three replicates for the indicated template total RNA amounts (NTC, no template control) are provided. ∆Cq is the difference between the Cq of the RT-qPCR with the indicated polymerase and control OneTube RT-PCR-Mix for each template dilution. (A) The real-time amplifications of GAPDH mRNA fragment with EvaGreen intercalating dye were carried out using 10-fold dilutions of the human total RNA from 1 ng to 0.01 ng as a template, or no template control (NTC). (B) The TaqMan prob RT-qPCR assays of SARS-CoV-2 viral RNA were carried out with 10, 50, and 250 times diluted RNA isolated from SARS-CoV-2-positive nasopharyngeal swabs as a template or no template control (NTC).

Article Snippet: The D732N mutant of Taq polymerase was commercialized by DNA Polymerase Technology, Inc. as the OmniTaq2 DNA polymerase.

Techniques: Comparison, Standard Deviation, Control, Isolation

Journal: Genetics and Molecular Research

Article Title: Regulation of ATG6/Beclin-1 homologs by abiotic stresses and hormones in rice (Oryza sativa L.)

doi: 10.4238/2012.august.17.3

Figure Lengend Snippet: Figure 6. Expression profile of OsATG6 genes under different abiotic stresses based on semi-quantitative real-time polymerase chain reaction.

Article Snippet: PCR was performed with 25 (18S rRNA) or 30 (ATG6 genes) cycles (30 s at 94°C, 30 s at 63°C, and 20 s at 72°C) under the following conditions: 0.5 μL RT product was amplified in a 20-μL volume containing 2 μL 10X PCR buffer with MgCl2, 0.25 μL 10 mM dNTPs, and 0.5 μL Taq polymerase (Tiangen, Beijing).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury

doi: 10.1371/journal.pone.0191853

Figure Lengend Snippet: Primer sequences for RT-PCR.

Article Snippet: To detect the Ezh2 floxed gene, one μl of genomic DNA from a mouse tail and primers for Ezh2 ( F: CTGCTCTGAATGGCAACTCC ; R: TTATTCATAGAGCCACCTGG ) were added to a mixture of solution containing Apex TaqDNA Polymerase (Cat. No. 42–409), Apex buffer, and MgCl 2 from Genesee Scientific (San Diego, CA), and dNTP (Cat. No. 10297–018; Invitrogen).

Techniques:

Journal: PLoS ONE

Article Title: Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury

doi: 10.1371/journal.pone.0191853

Figure Lengend Snippet: (A) PCR genotyping of Ezh2 and Cre genes. (B) Representative result of Western blot of Ezh2 expression in RGCs purified from P0 WT and mKO mice. GAPDH was used as a loading control. A strongly reduced level of Ezh2 was found in mKO RGCs as compared to WT RGCs. ( C) Epifluorescence images of retinal sections taken from P0 WT and mKO mice that were immunolabeled for H3K27me3 (red) and nuclear marker 4’,6-Diamidino-2-Phenylindole (DAPI; blue). Note the higher level of H3K27me3 signals in the mKO retina compared to WT retina. Scale bar: 50 μm.

Article Snippet: To detect the Ezh2 floxed gene, one μl of genomic DNA from a mouse tail and primers for Ezh2 ( F: CTGCTCTGAATGGCAACTCC ; R: TTATTCATAGAGCCACCTGG ) were added to a mixture of solution containing Apex TaqDNA Polymerase (Cat. No. 42–409), Apex buffer, and MgCl 2 from Genesee Scientific (San Diego, CA), and dNTP (Cat. No. 10297–018; Invitrogen).

Techniques: Western Blot, Expressing, Purification, Control, Immunolabeling, Marker

Journal: PLoS ONE

Article Title: Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury

doi: 10.1371/journal.pone.0191853

Figure Lengend Snippet: (A) Volcano plot showing fold changes (fc) of all genes detected from RGCs of mKO mice against control mice. Statistical significance (green dots) was defined as P <0.05 with a fold change ≥ +1.5 or ≤ -1.5 when compared to WT; non-significant changes (orange dots) fulfill either one or none of these two criteria. 997 genes were found with fc ≥ +1.5 and 1,220 genes with fc ≤ -1.5. Arrow points to the Ezh2 site. (B,C ) Pie charts represent depicted GO terms for upregulated (B) and downregulated (C) genes with │fc│ ≥ 1.5. No GO term in either up- or downregulated gene group were found to be specifically related to eye development. ( D) RT-PCR verification of mRNA levels of retinal related genes in RGCs purified from P5 floxed littermate control (white bar; n = 4) and mKO (black bar; n = 7) mouse pups. CRAL: Cellular retinaldehyde binding protein ; Tuj1: βIII-tubulin (Tubb3) ; Brn: Brn3a (Pou4f1) ; Rho: Rhodopsin ; Rec: Recoverin (* P < 0 . 05 by one-way ANOVA ).

Article Snippet: To detect the Ezh2 floxed gene, one μl of genomic DNA from a mouse tail and primers for Ezh2 ( F: CTGCTCTGAATGGCAACTCC ; R: TTATTCATAGAGCCACCTGG ) were added to a mixture of solution containing Apex TaqDNA Polymerase (Cat. No. 42–409), Apex buffer, and MgCl 2 from Genesee Scientific (San Diego, CA), and dNTP (Cat. No. 10297–018; Invitrogen).

Techniques: Control, Reverse Transcription Polymerase Chain Reaction, Purification, Binding Assay