sybr green power pcr master mix  (Thermo Fisher)


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    SYBR Green PCR Master Mix
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    Alternative Product Try PowerUp SYBR Green Master Mix our newest high performance SYBR dye based master mix for superior performance at a very competitive price With PowerUp SYBR Green Master Mix we ve taken the best of SYBR Green PCR Master Mix and added additional capabilities for your gene expression analysis Everything you need for SYBR Green dye based PCR amplification and detection in a convenient single tube format Applied Biosystems SYBR Green PCR Master Mix combines SYBR Green I dye AmpliTaq Gold DNA Polymerase dNTPs with dUTP Passive Reference 1 and optimized buffer in the convenience of a single vial • Premixed components stored at 2 8°C significantly reduce assay setup time• SYBR Green I dye detects double stranded DNA so specific probes are not required• AmpliTaq Gold DNA polymerase minimizes nonspecific product formation to achieve superior performance• dUTP significantly reduces carryover contamination when used in conjunction with uracil DNA glycosylase• Proprietary buffer enhancements ensure performance and reliabilityMaximum Flexibility and ConvenienceApplied Biosystems SYBR Green PCR Master Mix provides maximum flexibility at reduced cost because no target specific TaqMan probes are required SYBR Green I dye is a double stranded DNA binding dye that detects any double stranded DNA generated during PCR The hot start enzyme AmpliTaq Gold DNA Polymerase minimizes nonspecific product formation including primer dimers yielding superior performance and sensitivity Passive Internal Reference 1 is provided to normalize non PCR related fluorescence fluctuations This minimizes well to well variability that can result from a variety of causes such as pipetting error or sample evaporation SYBR Green I dye is ideal for target identification screening assays or when a limited number of assays is needed
    Catalog Number:
    4309155
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    Applications:
    Enzymes & Master Mixes for Real-Time PCR|PCR & Real-Time PCR|Real Time PCR (qPCR)|Real Time PCR-Based Gene Expression Profiling|Gene Expression Analysis & Genotyping
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    Structured Review

    Thermo Fisher sybr green power pcr master mix
    Alternative Product Try PowerUp SYBR Green Master Mix our newest high performance SYBR dye based master mix for superior performance at a very competitive price With PowerUp SYBR Green Master Mix we ve taken the best of SYBR Green PCR Master Mix and added additional capabilities for your gene expression analysis Everything you need for SYBR Green dye based PCR amplification and detection in a convenient single tube format Applied Biosystems SYBR Green PCR Master Mix combines SYBR Green I dye AmpliTaq Gold DNA Polymerase dNTPs with dUTP Passive Reference 1 and optimized buffer in the convenience of a single vial • Premixed components stored at 2 8°C significantly reduce assay setup time• SYBR Green I dye detects double stranded DNA so specific probes are not required• AmpliTaq Gold DNA polymerase minimizes nonspecific product formation to achieve superior performance• dUTP significantly reduces carryover contamination when used in conjunction with uracil DNA glycosylase• Proprietary buffer enhancements ensure performance and reliabilityMaximum Flexibility and ConvenienceApplied Biosystems SYBR Green PCR Master Mix provides maximum flexibility at reduced cost because no target specific TaqMan probes are required SYBR Green I dye is a double stranded DNA binding dye that detects any double stranded DNA generated during PCR The hot start enzyme AmpliTaq Gold DNA Polymerase minimizes nonspecific product formation including primer dimers yielding superior performance and sensitivity Passive Internal Reference 1 is provided to normalize non PCR related fluorescence fluctuations This minimizes well to well variability that can result from a variety of causes such as pipetting error or sample evaporation SYBR Green I dye is ideal for target identification screening assays or when a limited number of assays is needed
    https://www.bioz.com/result/sybr green power pcr master mix/product/Thermo Fisher
    Average 99 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    sybr green power pcr master mix - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed using SYBR Green PCR Master Mix (#4309155, Applied Biosystems, Foster City, CA) with 5 μl of immunoprecipitated material, 0.2 μM of primer [HIV-1 LTR (−69 − +175) Forward 5′-CTGGGCGGGACTGGGGAG-3′ and Reverse 5′-TCACACAACAGACGGGCACAC-3′]. .. The antibodies used for immunoprecipitation were as follows: total RNAP II CTD (ab817, Abcam), pS10-H3 (Novex, Life Technologies), BRCA1 (sc-642, Santa Cruz), p-BRCA1 S1423 (sc-101647, Santa, Cruz), Sp1 (5931, Cell Signaling), (IgG (sc-2027, Santa Cruz) or V5 (AbD Serotec, Oxford, UK).

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix Applied Biosystems). ..

    Article Title: The mouse complement regulator CD59b is significantly expressed only in testis and plays roles in sperm acrosome activation and motility
    Article Snippet: .. Quantitative PCR was performed on ABI PRISM 7000 using either TaqMan Universal PCR Master Mix or SYBR Green PCR Master Mix according to the manufacturer's instructions (Applied Biosystems) with 50 cycles of amplification. .. 2.5 Preparation of tissue lysates To obtain tissue lysates, freshly harvested mouse organs were immediately chilled and homogenized with ice-cold lysis buffer (PBS containing 2% NP40, 1 mM phenylmethylsulfonyl fluoride, 10 mM EDTA, 1 μg/ml leupeptin, and 1 μg/ml pepstatin; 1 g of tissue/1.5 ml of buffer) and incubated for 60 min on ice.

    Article Title: Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix (Thermo, F-415XL) on Applied Biosystems 7300 Fast Real-Time PCR System. ..

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology
    Article Snippet: .. 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix. .. For the SYBR green PCR assay, the 20 μl PCR mix reaction mix was comprised of 5 μl of cDNA and 15 μl of master mix prepared using 1 μl of the 10 pmol forward and reverse primers, 10 μl SYBR® Green I and 3 μl of nuclease free water.

    Amplification:

    Article Title: The mouse complement regulator CD59b is significantly expressed only in testis and plays roles in sperm acrosome activation and motility
    Article Snippet: .. Quantitative PCR was performed on ABI PRISM 7000 using either TaqMan Universal PCR Master Mix or SYBR Green PCR Master Mix according to the manufacturer's instructions (Applied Biosystems) with 50 cycles of amplification. .. 2.5 Preparation of tissue lysates To obtain tissue lysates, freshly harvested mouse organs were immediately chilled and homogenized with ice-cold lysis buffer (PBS containing 2% NP40, 1 mM phenylmethylsulfonyl fluoride, 10 mM EDTA, 1 μg/ml leupeptin, and 1 μg/ml pepstatin; 1 g of tissue/1.5 ml of buffer) and incubated for 60 min on ice.

    Article Title: RNase L restricts the mobility of engineered retrotransposons in cultured human cells
    Article Snippet: .. The resultant cDNA was amplified using Sybr Green PCR master mix (Gibco/Life Technologies) on a StepOnePlus system according to manufacturer’s protocol. ..

    In Vitro:

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology
    Article Snippet: .. 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix. .. For the SYBR green PCR assay, the 20 μl PCR mix reaction mix was comprised of 5 μl of cDNA and 15 μl of master mix prepared using 1 μl of the 10 pmol forward and reverse primers, 10 μl SYBR® Green I and 3 μl of nuclease free water.

    Quantitative RT-PCR:

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology
    Article Snippet: .. 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix. .. For the SYBR green PCR assay, the 20 μl PCR mix reaction mix was comprised of 5 μl of cDNA and 15 μl of master mix prepared using 1 μl of the 10 pmol forward and reverse primers, 10 μl SYBR® Green I and 3 μl of nuclease free water.

    SYBR Green Assay:

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed using SYBR Green PCR Master Mix (#4309155, Applied Biosystems, Foster City, CA) with 5 μl of immunoprecipitated material, 0.2 μM of primer [HIV-1 LTR (−69 − +175) Forward 5′-CTGGGCGGGACTGGGGAG-3′ and Reverse 5′-TCACACAACAGACGGGCACAC-3′]. .. The antibodies used for immunoprecipitation were as follows: total RNAP II CTD (ab817, Abcam), pS10-H3 (Novex, Life Technologies), BRCA1 (sc-642, Santa Cruz), p-BRCA1 S1423 (sc-101647, Santa, Cruz), Sp1 (5931, Cell Signaling), (IgG (sc-2027, Santa Cruz) or V5 (AbD Serotec, Oxford, UK).

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix Applied Biosystems). ..

    Article Title: The mouse complement regulator CD59b is significantly expressed only in testis and plays roles in sperm acrosome activation and motility
    Article Snippet: .. Quantitative PCR was performed on ABI PRISM 7000 using either TaqMan Universal PCR Master Mix or SYBR Green PCR Master Mix according to the manufacturer's instructions (Applied Biosystems) with 50 cycles of amplification. .. 2.5 Preparation of tissue lysates To obtain tissue lysates, freshly harvested mouse organs were immediately chilled and homogenized with ice-cold lysis buffer (PBS containing 2% NP40, 1 mM phenylmethylsulfonyl fluoride, 10 mM EDTA, 1 μg/ml leupeptin, and 1 μg/ml pepstatin; 1 g of tissue/1.5 ml of buffer) and incubated for 60 min on ice.

    Article Title: RNase L restricts the mobility of engineered retrotransposons in cultured human cells
    Article Snippet: .. The resultant cDNA was amplified using Sybr Green PCR master mix (Gibco/Life Technologies) on a StepOnePlus system according to manufacturer’s protocol. ..

    Article Title: Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix (Thermo, F-415XL) on Applied Biosystems 7300 Fast Real-Time PCR System. ..

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology
    Article Snippet: .. 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix. .. For the SYBR green PCR assay, the 20 μl PCR mix reaction mix was comprised of 5 μl of cDNA and 15 μl of master mix prepared using 1 μl of the 10 pmol forward and reverse primers, 10 μl SYBR® Green I and 3 μl of nuclease free water.

    Article Title: A Myc-microRNA network promotes exit from quiescence by suppressing the interferon response and cell-cycle arrest genes
    Article Snippet: .. PCR was performed using the SYBR GREEN PCR Master Mix from Applied Biosystems. .. The target-gene messenger RNA (mRNA) expression was normalized to the expression of GAPDH, and relative mRNA fold changes were calculated by the ΔΔCt method.

    Article Title: hCAF1/CNOT7 regulates interferon signalling by targeting STAT1
    Article Snippet: .. After DNA purification, qPCRs were performed in triplicate in a 96-well optical reaction plate using SYBR Green PCR Master Mix (Life Technology). .. The –ΔΔCt values for each locus were calculated with respect to the ChIP input DNA, normalized to a reference locus (3′ downstream region of the GAPDH gene).

    Immunoprecipitation:

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed using SYBR Green PCR Master Mix (#4309155, Applied Biosystems, Foster City, CA) with 5 μl of immunoprecipitated material, 0.2 μM of primer [HIV-1 LTR (−69 − +175) Forward 5′-CTGGGCGGGACTGGGGAG-3′ and Reverse 5′-TCACACAACAGACGGGCACAC-3′]. .. The antibodies used for immunoprecipitation were as follows: total RNAP II CTD (ab817, Abcam), pS10-H3 (Novex, Life Technologies), BRCA1 (sc-642, Santa Cruz), p-BRCA1 S1423 (sc-101647, Santa, Cruz), Sp1 (5931, Cell Signaling), (IgG (sc-2027, Santa Cruz) or V5 (AbD Serotec, Oxford, UK).

    DNA Purification:

    Article Title: hCAF1/CNOT7 regulates interferon signalling by targeting STAT1
    Article Snippet: .. After DNA purification, qPCRs were performed in triplicate in a 96-well optical reaction plate using SYBR Green PCR Master Mix (Life Technology). .. The –ΔΔCt values for each locus were calculated with respect to the ChIP input DNA, normalized to a reference locus (3′ downstream region of the GAPDH gene).

    Polymerase Chain Reaction:

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed using SYBR Green PCR Master Mix (#4309155, Applied Biosystems, Foster City, CA) with 5 μl of immunoprecipitated material, 0.2 μM of primer [HIV-1 LTR (−69 − +175) Forward 5′-CTGGGCGGGACTGGGGAG-3′ and Reverse 5′-TCACACAACAGACGGGCACAC-3′]. .. The antibodies used for immunoprecipitation were as follows: total RNAP II CTD (ab817, Abcam), pS10-H3 (Novex, Life Technologies), BRCA1 (sc-642, Santa Cruz), p-BRCA1 S1423 (sc-101647, Santa, Cruz), Sp1 (5931, Cell Signaling), (IgG (sc-2027, Santa Cruz) or V5 (AbD Serotec, Oxford, UK).

    Article Title: BRCA1 functions as a novel transcriptional cofactor in HIV-1 infection
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix Applied Biosystems). ..

    Article Title: The mouse complement regulator CD59b is significantly expressed only in testis and plays roles in sperm acrosome activation and motility
    Article Snippet: .. Quantitative PCR was performed on ABI PRISM 7000 using either TaqMan Universal PCR Master Mix or SYBR Green PCR Master Mix according to the manufacturer's instructions (Applied Biosystems) with 50 cycles of amplification. .. 2.5 Preparation of tissue lysates To obtain tissue lysates, freshly harvested mouse organs were immediately chilled and homogenized with ice-cold lysis buffer (PBS containing 2% NP40, 1 mM phenylmethylsulfonyl fluoride, 10 mM EDTA, 1 μg/ml leupeptin, and 1 μg/ml pepstatin; 1 g of tissue/1.5 ml of buffer) and incubated for 60 min on ice.

    Article Title: RNase L restricts the mobility of engineered retrotransposons in cultured human cells
    Article Snippet: .. The resultant cDNA was amplified using Sybr Green PCR master mix (Gibco/Life Technologies) on a StepOnePlus system according to manufacturer’s protocol. ..

    Article Title: Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways
    Article Snippet: .. Quantitative PCR was performed with SYBR Green PCR Master Mix (Thermo, F-415XL) on Applied Biosystems 7300 Fast Real-Time PCR System. ..

    Article Title: A Myc-microRNA network promotes exit from quiescence by suppressing the interferon response and cell-cycle arrest genes
    Article Snippet: .. PCR was performed using the SYBR GREEN PCR Master Mix from Applied Biosystems. .. The target-gene messenger RNA (mRNA) expression was normalized to the expression of GAPDH, and relative mRNA fold changes were calculated by the ΔΔCt method.

    Article Title: hCAF1/CNOT7 regulates interferon signalling by targeting STAT1
    Article Snippet: .. After DNA purification, qPCRs were performed in triplicate in a 96-well optical reaction plate using SYBR Green PCR Master Mix (Life Technology). .. The –ΔΔCt values for each locus were calculated with respect to the ChIP input DNA, normalized to a reference locus (3′ downstream region of the GAPDH gene).

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    Thermo Fisher power sybr green pcr master mix
    Hypermutation of the FeLV-B genome and APOBEC3G mRNA expression in human cells. FeLV genomic sequences (segments of Gag and Pol) were cloned by <t>PCR</t> from infected human cells, and individual templates were sequenced and compared to the reference input virus (pFGB clone). APOBEC3G mRNA expression was determined in the same cells (prior to infection) by quantitative real-time PCR (with <t>SYBR</t> green). (A) Representative plots of hypermutation visualized by the online HYPERMUT program ( www.hiv.lanl.gov/content/sequence/HYPERMUT/hypermut.html ), where sequence changes relative to the reference FeLV-B genome are color coded (red, GG→AG; cyan, GA→AA; green, GC→AC; magenta, GT→AT; black, non-G→A). (B) X/Y plots of G→A mutation (per kilobase) against APOBEC3G mRNA levels (where the level in HEK293 cells is taken as 1) with cell lines sorted according to FeLV restriction phenotype. Results for LCLs, which are discordant by virtue of their low levels of infectious virion release ( Table 2 ) but postinfection accumulation of proviral DNA ( Fig. 3 ), are enclosed by a dashed oval. (C) Percentage of G→A mutations that conform to the A3G signature ( 42 ) for all cell lines in which significant levels of mutations were detected. Blue-gray bars, hematopoietic cells; yellow bars, nonhematopoietic cells. (D) Relative levels of APOBEC3G mRNA (on a log 10 scale, with the level in HEK293 cells taken as 1) for all the cell lines tested, sorted into hematopoietic (blue circles) and nonhematopoietic (green circles) cell lines. Nonpermissive cells (nonspreading, with low virion release) are represented by black circles.
    Power Sybr Green Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8809 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/power sybr green pcr master mix/product/Thermo Fisher
    Average 99 stars, based on 8809 article reviews
    Price from $9.99 to $1999.99
    power sybr green pcr master mix - by Bioz Stars, 2020-09
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    99
    Thermo Fisher power sybr green rna to ct rt qpcr kit
    COS hiPSC cohort reprogramming and differentiation. a Validated hiPSCs (from 14 individuals with childhood-onset-schizophrenia (COS) and 12 unrelated healthy controls) and NPCs (12 COS; 12 control individuals) yielded 94 <t>RNA-Seq</t> samples (11 COS; 11 control individuals). b Schematic illustration of the reprogramming and differentiation process, noting the yield at each stage. c Sex breakdown of the COS-control cohort. d Breakdown of SZ-associated copy number variants in the 11 COS patients with RNA-Seq data. e Representative <t>qPCR</t> validation of NANOG, NESTIN , and SYN1 expression in hiPSCs (white bar), NPCs (light gray) and 6-week-old neurons (dark gray) from three individuals. f FACS analysis for pluripotency markers TRA-1-60 (left) and SSEA4 (right) in representative control (blue, n = 17) and COS (red, n = 16) hiPSCs. g FACS analysis for NPC markers SOX2 (left) and NESTIN (right) in control (blue, n = 34) and COS (red, n = 37) NPCs. h Representative images of NPCs (left) and 6-week-old forebrain neurons (right) from control (top) and COS (bottom). NPCs stained with SOX2 (red) and NESTIN (green); neurons stained with MAP2 (red). DAPI-stained nuclei (blue). Scale bar=50 μm. i Computational workflow showing quality control, integration with external data sets, computational deconvolution with Cibersort, decomposition multiple sources of expression variation with variancePartition, coexpression analysis with WGCNA, differential expression and concordance analysis
    Power Sybr Green Rna To Ct Rt Qpcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/power sybr green rna to ct rt qpcr kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    power sybr green rna to ct rt qpcr kit - by Bioz Stars, 2020-09
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    Hypermutation of the FeLV-B genome and APOBEC3G mRNA expression in human cells. FeLV genomic sequences (segments of Gag and Pol) were cloned by PCR from infected human cells, and individual templates were sequenced and compared to the reference input virus (pFGB clone). APOBEC3G mRNA expression was determined in the same cells (prior to infection) by quantitative real-time PCR (with SYBR green). (A) Representative plots of hypermutation visualized by the online HYPERMUT program ( www.hiv.lanl.gov/content/sequence/HYPERMUT/hypermut.html ), where sequence changes relative to the reference FeLV-B genome are color coded (red, GG→AG; cyan, GA→AA; green, GC→AC; magenta, GT→AT; black, non-G→A). (B) X/Y plots of G→A mutation (per kilobase) against APOBEC3G mRNA levels (where the level in HEK293 cells is taken as 1) with cell lines sorted according to FeLV restriction phenotype. Results for LCLs, which are discordant by virtue of their low levels of infectious virion release ( Table 2 ) but postinfection accumulation of proviral DNA ( Fig. 3 ), are enclosed by a dashed oval. (C) Percentage of G→A mutations that conform to the A3G signature ( 42 ) for all cell lines in which significant levels of mutations were detected. Blue-gray bars, hematopoietic cells; yellow bars, nonhematopoietic cells. (D) Relative levels of APOBEC3G mRNA (on a log 10 scale, with the level in HEK293 cells taken as 1) for all the cell lines tested, sorted into hematopoietic (blue circles) and nonhematopoietic (green circles) cell lines. Nonpermissive cells (nonspreading, with low virion release) are represented by black circles.

    Journal: Journal of Virology

    Article Title: Barriers to Infection of Human Cells by Feline Leukemia Virus: Insights into Resistance to Zoonosis

    doi: 10.1128/JVI.02119-16

    Figure Lengend Snippet: Hypermutation of the FeLV-B genome and APOBEC3G mRNA expression in human cells. FeLV genomic sequences (segments of Gag and Pol) were cloned by PCR from infected human cells, and individual templates were sequenced and compared to the reference input virus (pFGB clone). APOBEC3G mRNA expression was determined in the same cells (prior to infection) by quantitative real-time PCR (with SYBR green). (A) Representative plots of hypermutation visualized by the online HYPERMUT program ( www.hiv.lanl.gov/content/sequence/HYPERMUT/hypermut.html ), where sequence changes relative to the reference FeLV-B genome are color coded (red, GG→AG; cyan, GA→AA; green, GC→AC; magenta, GT→AT; black, non-G→A). (B) X/Y plots of G→A mutation (per kilobase) against APOBEC3G mRNA levels (where the level in HEK293 cells is taken as 1) with cell lines sorted according to FeLV restriction phenotype. Results for LCLs, which are discordant by virtue of their low levels of infectious virion release ( Table 2 ) but postinfection accumulation of proviral DNA ( Fig. 3 ), are enclosed by a dashed oval. (C) Percentage of G→A mutations that conform to the A3G signature ( 42 ) for all cell lines in which significant levels of mutations were detected. Blue-gray bars, hematopoietic cells; yellow bars, nonhematopoietic cells. (D) Relative levels of APOBEC3G mRNA (on a log 10 scale, with the level in HEK293 cells taken as 1) for all the cell lines tested, sorted into hematopoietic (blue circles) and nonhematopoietic (green circles) cell lines. Nonpermissive cells (nonspreading, with low virion release) are represented by black circles.

    Article Snippet: Aliquots (12.5 ng) of cDNA were amplified in triplicate using Power SYBR green PCR master mix (Thermo Fisher Scientific) and Qiagen QuantiTect primer assays for human hypoxanthine phosphoribosyltransferase (HPRT) (QT00059066), APOBEC3G (QT00070770), SLC20A1 (PIT-1) (QT00028763), or SLC19A2 (THTR1) (QT00007847) on an ABI 7500 real-time PCR system.

    Techniques: Expressing, Genomic Sequencing, Clone Assay, Polymerase Chain Reaction, Infection, Real-time Polymerase Chain Reaction, SYBR Green Assay, Sequencing, Mutagenesis

    The thymus produces a significant number of Pth transcripts that increase after TPTX. (A) The thymus produced Pth transcripts. Mice in which serum PTH levels recovered (n = 10) were euthanized, and the heart, lung, liver, spleen, kidney, pancreas, testis or ovary, hypothalamus, pituitary gland, thymus, and adrenal glands were harvested. RNA qRT-PCR was performed on the tissue samples using SYBR Green/Taqman Master Mix with custom primers for the mouse Pth transcript and a commercial probe-set for 18S rRNA (endogenous control). Expression data were normalized to 18S rRNA, and fold changes of expression against arbitrary numbers were calculated using the 2 − ΔΔ Ct ). (B) Pth transcripts increased after TPTX. Thymi from unoperated mice (control) were harvested, and the same procedures were performed as in (A). qRT-PCR was performed as described with a commercial probe (gray) and custom probes (black). Student t test was performed. ** P

    Journal: Endocrinology

    Article Title: Thymic PTH Increases After Thyroparathyroidectomy in C57BL/KaLwRij Mice

    doi: 10.1210/en.2017-03083

    Figure Lengend Snippet: The thymus produces a significant number of Pth transcripts that increase after TPTX. (A) The thymus produced Pth transcripts. Mice in which serum PTH levels recovered (n = 10) were euthanized, and the heart, lung, liver, spleen, kidney, pancreas, testis or ovary, hypothalamus, pituitary gland, thymus, and adrenal glands were harvested. RNA qRT-PCR was performed on the tissue samples using SYBR Green/Taqman Master Mix with custom primers for the mouse Pth transcript and a commercial probe-set for 18S rRNA (endogenous control). Expression data were normalized to 18S rRNA, and fold changes of expression against arbitrary numbers were calculated using the 2 − ΔΔ Ct ). (B) Pth transcripts increased after TPTX. Thymi from unoperated mice (control) were harvested, and the same procedures were performed as in (A). qRT-PCR was performed as described with a commercial probe (gray) and custom probes (black). Student t test was performed. ** P

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) was performed with the Power SYBR Green PCR Master Mix on the Step-One-Plus system (Thermo Fisher Scientific).

    Techniques: Produced, Mouse Assay, Quantitative RT-PCR, SYBR Green Assay, Expressing

    Accumulation of immunostimulatory 5′-ppp-vtRNAs during lytic reactivation. a Predicted secondary structure of vtRNAs generated by RNAfold. b SYBR-Gold staining of in vitro transcribed vtRNAs with or without CIP treatment. c HCT116 ISG54-luciferase reporter cells were transfected with 100 ng in vitro transcribed vtRNAs with or without CIP treatment. Cells were harvested 24 h posttransfection and subjected to luciferase assay. Mock indicated cells without RNA transfection and was set as 1. d BC-3 cells were reactivated for 3 days and expression of DUSP11 was quantified by RT-qPCR. L latency, D1–D3 lytic reactivation for 1 day to 3 days. The DUSP11 expression was normalized to the level of 18S rRNA and L was set as 1. e Cell lysates were prepared from BC-3 cells described in ( d ) and DUSP11 protein levels were monitored by Western blot. GAPDH was run as a loading control. f Latent and lytic BC-3 cells were subjected to RNAP II ChIP-qPCR analysis. Signals were normalized to input. g ). h HCT116 ISG54-luciferase reporter cells were transfected with vtRNA or U1 RNA isolated by antisense oligonucleotide affinity selection from either latent or lytic BC-3 cells. Cells were harvested 12 h posttransfection and subjected to luciferase assay. Mock indicated cells without RNA transfection and was set as 1. Error bars in all panels represent mean ± SD from three independent experiments. p Values were determined by the Student’s t test, * p

    Journal: Nature Communications

    Article Title: RIG-I like receptor sensing of host RNAs facilitates the cell-intrinsic immune response to KSHV infection

    doi: 10.1038/s41467-018-07314-7

    Figure Lengend Snippet: Accumulation of immunostimulatory 5′-ppp-vtRNAs during lytic reactivation. a Predicted secondary structure of vtRNAs generated by RNAfold. b SYBR-Gold staining of in vitro transcribed vtRNAs with or without CIP treatment. c HCT116 ISG54-luciferase reporter cells were transfected with 100 ng in vitro transcribed vtRNAs with or without CIP treatment. Cells were harvested 24 h posttransfection and subjected to luciferase assay. Mock indicated cells without RNA transfection and was set as 1. d BC-3 cells were reactivated for 3 days and expression of DUSP11 was quantified by RT-qPCR. L latency, D1–D3 lytic reactivation for 1 day to 3 days. The DUSP11 expression was normalized to the level of 18S rRNA and L was set as 1. e Cell lysates were prepared from BC-3 cells described in ( d ) and DUSP11 protein levels were monitored by Western blot. GAPDH was run as a loading control. f Latent and lytic BC-3 cells were subjected to RNAP II ChIP-qPCR analysis. Signals were normalized to input. g ). h HCT116 ISG54-luciferase reporter cells were transfected with vtRNA or U1 RNA isolated by antisense oligonucleotide affinity selection from either latent or lytic BC-3 cells. Cells were harvested 12 h posttransfection and subjected to luciferase assay. Mock indicated cells without RNA transfection and was set as 1. Error bars in all panels represent mean ± SD from three independent experiments. p Values were determined by the Student’s t test, * p

    Article Snippet: RNA was DNase I (NEB) treated at 37 °C for 20 min, and inactivated with EDTA at 70 °C for 10 min. cDNA was synthesized from DNase-treated RNA with random 9-mer (Integrated DNA Technologies) and M-MLV RT (Promega). qPCR was performed using the PowerUp SYBR Green qPCR kit (Thermo Scientific) with appropriate primers (Supplementary Table ).

    Techniques: Generated, Staining, In Vitro, Luciferase, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Isolation, Selection

    COS hiPSC cohort reprogramming and differentiation. a Validated hiPSCs (from 14 individuals with childhood-onset-schizophrenia (COS) and 12 unrelated healthy controls) and NPCs (12 COS; 12 control individuals) yielded 94 RNA-Seq samples (11 COS; 11 control individuals). b Schematic illustration of the reprogramming and differentiation process, noting the yield at each stage. c Sex breakdown of the COS-control cohort. d Breakdown of SZ-associated copy number variants in the 11 COS patients with RNA-Seq data. e Representative qPCR validation of NANOG, NESTIN , and SYN1 expression in hiPSCs (white bar), NPCs (light gray) and 6-week-old neurons (dark gray) from three individuals. f FACS analysis for pluripotency markers TRA-1-60 (left) and SSEA4 (right) in representative control (blue, n = 17) and COS (red, n = 16) hiPSCs. g FACS analysis for NPC markers SOX2 (left) and NESTIN (right) in control (blue, n = 34) and COS (red, n = 37) NPCs. h Representative images of NPCs (left) and 6-week-old forebrain neurons (right) from control (top) and COS (bottom). NPCs stained with SOX2 (red) and NESTIN (green); neurons stained with MAP2 (red). DAPI-stained nuclei (blue). Scale bar=50 μm. i Computational workflow showing quality control, integration with external data sets, computational deconvolution with Cibersort, decomposition multiple sources of expression variation with variancePartition, coexpression analysis with WGCNA, differential expression and concordance analysis

    Journal: Nature Communications

    Article Title: Transcriptional signatures of schizophrenia in hiPSC-derived NPCs and neurons are concordant with post-mortem adult brains

    doi: 10.1038/s41467-017-02330-5

    Figure Lengend Snippet: COS hiPSC cohort reprogramming and differentiation. a Validated hiPSCs (from 14 individuals with childhood-onset-schizophrenia (COS) and 12 unrelated healthy controls) and NPCs (12 COS; 12 control individuals) yielded 94 RNA-Seq samples (11 COS; 11 control individuals). b Schematic illustration of the reprogramming and differentiation process, noting the yield at each stage. c Sex breakdown of the COS-control cohort. d Breakdown of SZ-associated copy number variants in the 11 COS patients with RNA-Seq data. e Representative qPCR validation of NANOG, NESTIN , and SYN1 expression in hiPSCs (white bar), NPCs (light gray) and 6-week-old neurons (dark gray) from three individuals. f FACS analysis for pluripotency markers TRA-1-60 (left) and SSEA4 (right) in representative control (blue, n = 17) and COS (red, n = 16) hiPSCs. g FACS analysis for NPC markers SOX2 (left) and NESTIN (right) in control (blue, n = 34) and COS (red, n = 37) NPCs. h Representative images of NPCs (left) and 6-week-old forebrain neurons (right) from control (top) and COS (bottom). NPCs stained with SOX2 (red) and NESTIN (green); neurons stained with MAP2 (red). DAPI-stained nuclei (blue). Scale bar=50 μm. i Computational workflow showing quality control, integration with external data sets, computational deconvolution with Cibersort, decomposition multiple sources of expression variation with variancePartition, coexpression analysis with WGCNA, differential expression and concordance analysis

    Article Snippet: Transcript analysis was carried out using a QuantStudio™ 7 Flex Real-Time PCR System using the Power SYBR green RNA-to-Ct RT-qPCR kit for primers (all ThermoFisher Scientific).

    Techniques: RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Expressing, FACS, Staining