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Roche two step nested long range pcr protocol
Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension <t>PCR</t> showing large-scale <t>mtDNA</t> deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p
Two Step Nested Long Range Pcr Protocol, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/two step nested long range pcr protocol/product/Roche
Average 85 stars, based on 1 article reviews
Price from $9.99 to $1999.99
two step nested long range pcr protocol - by Bioz Stars, 2020-08
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1) Product Images from "TOM40 Mediates Mitochondrial Dysfunction Induced by ?-Synuclein Accumulation in Parkinson's Disease"

Article Title: TOM40 Mediates Mitochondrial Dysfunction Induced by ?-Synuclein Accumulation in Parkinson's Disease

Journal: PLoS ONE

doi: 10.1371/journal.pone.0062277

Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension PCR showing large-scale mtDNA deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p
Figure Legend Snippet: Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension PCR showing large-scale mtDNA deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p

Techniques Used: Over Expression, Transgenic Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Immunostaining, Labeling, Laser Capture Microdissection, Western Blot, Marker

Related Articles

Polymerase Chain Reaction:

Article Title: TOM40 Mediates Mitochondrial Dysfunction Induced by ?-Synuclein Accumulation in Parkinson's Disease
Article Snippet: .. Low-abundance mtDNA deletions were detected by a two-step nested long-range PCR protocol (Expand Long Range dNTPack®, Roche, Mannheim, Germany) to amplify a final ∼10 kb fragment of the deletion-prone “major arc” of the mitochondrial genome as we previously reported , . .. In brief, first round PCR reactions were done on 10 ng of template DNA under standard conditions and using 30 pmol each of forward (mtDNA nt 4796-4817) and reverse (mtDNA nt 16020-16001) primers.

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    Roche two step nested long range pcr protocol
    Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension <t>PCR</t> showing large-scale <t>mtDNA</t> deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p
    Two Step Nested Long Range Pcr Protocol, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/two step nested long range pcr protocol/product/Roche
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    two step nested long range pcr protocol - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

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    Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension PCR showing large-scale mtDNA deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p

    Journal: PLoS ONE

    Article Title: TOM40 Mediates Mitochondrial Dysfunction Induced by ?-Synuclein Accumulation in Parkinson's Disease

    doi: 10.1371/journal.pone.0062277

    Figure Lengend Snippet: Alpha-synuclein overexpression results in oxidative DNA damage, alterations in respiratory chain complex I and mitochondrial DNA deletions in transgenic mice brains. (A) Immuno-histochemical detection of 8-OHdG suggesting increased oxidative DNA damage in α-Syn tg mice brains. (B) 8-OHdG levels were quantified on whole brain in α-Syn tg and non-transgenic control mice by ELISA. Scale bar represents 25 µm. (C) Long extension PCR showing large-scale mtDNA deletions in α-Syn tg mice. (D) Quantitative determination of mtDNA deletions by real-time PCR showing increased mtDNA deletion levels in the brain of α-Syn tg mice compared to control animals. (E) Immunostaining for α-Syn (green signal) and NeuN (red signal; arrow marks the identical neuron in double labeling) used for Laser Capture Microdissection (LCM). Scale bar represents 250 µm in the upper panel and 25 µm in the close-up lower panel. (F) Quantification of mtDNA deletion levels by real-time PCR on individual neurons with either intense or negative α-Syn immunoreactivity after LCM. (G) Western blot analysis of mitochondrial OXPHOS in brain homogenates from α-Syn tg and wt mice. Lane 1 shows mitochondrial proteins standard marker. (H) Densitometric analysis of the levels of Complexes I, II, III and IV. *p

    Article Snippet: Low-abundance mtDNA deletions were detected by a two-step nested long-range PCR protocol (Expand Long Range dNTPack®, Roche, Mannheim, Germany) to amplify a final ∼10 kb fragment of the deletion-prone “major arc” of the mitochondrial genome as we previously reported , .

    Techniques: Over Expression, Transgenic Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Immunostaining, Labeling, Laser Capture Microdissection, Western Blot, Marker