dna binding agent  (Thermo Fisher)

Journal: PLoS ONE

Article Title: The Polymorphic AluYb8 Insertion in the MUTYH Gene is Associated with Reduced Type 1 Protein Expression and Reduced Mitochondrial DNA Content

doi: 10.1371/journal.pone.0070718

Figure Lengend Snippet: (A) Schematic representation of the genomic structure of MUTYH from exon 15 to 16 indicates the position of the AluYb8 insertion in intron 15, depicted as a deep blue arrow. Exons are shown by boxes. The genotyping primers are represented by black arrowheads, and the lengths of their PCR products are also depicted. (B) A schematic illustration of the alternative splicing pattern between exons 14 and 16 of the MUTYH gene. Scores (MaxEnt score) of splice sites for the three exons were recorded in red. An alternatively spliced cassette exon within intron 15 was inferred from aligning two ESTs (BM679345.1 and AW518294.1) with human genomic DNA, and the putative cassette exon in the variant intron 15 is shown. (C) Splicing assays were performed in healthy adults. A representative ethidium bromide-stained agarose gel separating the RT-PCR products spanning MUTYH exon 14 to exon 16 are shown. PCR products were confirmed by sequencing (the bottom of the agarose gel electrophoresis). (D) The constructed wild-type (wt) and mutant minigene plasmids were transfected into 293 T cells, respectively, total RNA was extracted, and splicing products were separated on a 2% agarose gel after RT-PCR analysis. PCR products were sequenced. Similar results were obtained in two independent experiments.

Article Snippet: PCR products were quantified using fluorescence measurements by the PicoGreen double-stranded DNA binding agent (Invitrogen).

Techniques: Variant Assay, Staining, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Sequencing, Construct, Mutagenesis, Transfection

Journal: PLoS ONE

Article Title: The Polymorphic AluYb8 Insertion in the MUTYH Gene is Associated with Reduced Type 1 Protein Expression and Reduced Mitochondrial DNA Content

doi: 10.1371/journal.pone.0070718

Figure Lengend Snippet: The long-range PCR was performed for both the mitochondrial DNA (A, open bars) and nuclear β-globin fragments (B, solid bars) among the healthy individuals aged 22 to 44 with different AluYb8MUTYH genotypes. The boxes cover the 25 th to 75 th percentiles, and the minimal and maximal values are shown by the ends of the bars. Relative amplification is presented relative to average of A/A group values, n = 18. P -values are indicated, multinomial logistic regression.

Article Snippet: PCR products were quantified using fluorescence measurements by the PicoGreen double-stranded DNA binding agent (Invitrogen).

Techniques: Amplification

Journal: Frontiers in Pediatrics

Article Title: Exposure to Morphine and Caffeine Induces Apoptosis and Mitochondrial Dysfunction in a Neonatal Rat Brain

doi: 10.3389/fped.2020.00593

Figure Lengend Snippet: DNA Damage: Representative fluorescence microscopy images of brain tissue sections of Control, MOR, CAFF, and M+C treated pups at PND 7, PND 14, and PND 28 and a graph of quantified 7AAD + cells. Scale bar = 100 μm. Values are expressed as mean ± S.E.M. * P < 0.001 vs. Control.

Article Snippet: The brain sections were incubated with fluorescent DNA binding agent, 7AAD (0.25 μg/mL; Thermo Fisher Scientific, Bartlett, IL) for 15 min at 4°C.

Techniques: Fluorescence, Microscopy, Control

Journal: Frontiers in Pediatrics

Article Title: Exposure to Morphine and Caffeine Induces Apoptosis and Mitochondrial Dysfunction in a Neonatal Rat Brain

doi: 10.3389/fped.2020.00593

Figure Lengend Snippet:

Article Snippet: The brain sections were incubated with fluorescent DNA binding agent, 7AAD (0.25 μg/mL; Thermo Fisher Scientific, Bartlett, IL) for 15 min at 4°C.

Techniques: Saline