moloney murine leukaemia virus reverse transcriptase  (Thermo Fisher)


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    Thermo Fisher moloney murine leukaemia virus reverse transcriptase
    Analysis of human class A scavenger receptor (hSR-A) III transgene RNA expression. (a) Northern blot of isolated macrophage (Mφ) populations. Total RNA was prepared from peritoneal cells isolated from transgenic line III.11 (Tg + ) mice and non-transgenic (Tg – ) littermates, either 1 or 4 days after injection of 1 ml of thioglycollate broth. Bone marrow-derived Mφs (BMDMs) were generated by the culture of bone marrow precursors in medium containing macrophage colony-stimulating factor (M-CSF) and total RNA isolated from adherent cells cultured for 6 days. Total RNA (5 µg) was denatured and separated on a 1% formaldehyde–agarose gel prior to capillary transfer to a nylon membrane. Blots were hybridized with [α- 32 P]dATP-labelled DNA probes for β-actin (a 2-kb fragment of human β-actin; Clontech) or hSR-A III transgene (a 1·2-kb fragment of type III hSR-A coding sequence generated by digestion of expression plasmid pcDNA 3 with Hin dIII and Xba I) and exposed to film. (b) Northern blot of adult tissues. Total tissue RNA (10 µg) prepared using organs collected from transgenic line III.4 (Tg + ) mice and non-transgenic (Tg – ) littermates, and from human monocyte-derived Mφ total RNA (10 µg), was denatured and separated on a 1% formaldehyde–agarose gel prior to transfer and hybridization as described in (a). The different sizes of SR-A hybridizing bands in the human and transgenic RNA samples are caused by the absence of 3′ untranslated sequences in the CD68 SR-A type III transgene. (c) Reverse transcription–polymerase chain reaction (RT–PCR) of adult tissues. Total tissue RNA from organs collected from transgenic line III.4 (Tg + ) mice was reverse transcribed in the presence (+) or absence (–) of <t>Moloney</t> murine leukemia virus reverse transcriptase. cDNA synthesized from 100 ng of total RNA served as a template for subsequent PCR using primers specific for hypoxanthine phosphoribosyl transferase (HPRT), macrosialin or the hSR-A III transgene. To enable a comparison of the relative levels of RNA for macrosialin and the hSR-A III transgene, PCR was performed with both oligonucleotide pairs in the same reaction tube. Primers for HPRT and macrosialin do not generate a detectable reaction product from contaminating genomic DNA (gDNA) under the conditions used, while the product corresponding to the amplification of the hSR-A III transgene genomic DNA is 83 bp larger owing to the presence of the first intron (IVS-1) sequence, which is spliced out during the processing of the primary RNA transcript. Reaction products were separated on a 1·2% agarose gel and visualized by ethidium bromide staining.
    Moloney Murine Leukaemia Virus Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/moloney murine leukaemia virus reverse transcriptase/product/Thermo Fisher
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    moloney murine leukaemia virus reverse transcriptase - by Bioz Stars, 2020-05
    93/100 stars

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    1) Product Images from "The use of human CD68 transcriptional regulatory sequences to direct high-level expression of class A scavenger receptor in macrophages in vitro and in vivo"

    Article Title: The use of human CD68 transcriptional regulatory sequences to direct high-level expression of class A scavenger receptor in macrophages in vitro and in vivo

    Journal: Immunology

    doi: 10.1046/j.1365-2567.2001.01256.x

    Analysis of human class A scavenger receptor (hSR-A) III transgene RNA expression. (a) Northern blot of isolated macrophage (Mφ) populations. Total RNA was prepared from peritoneal cells isolated from transgenic line III.11 (Tg + ) mice and non-transgenic (Tg – ) littermates, either 1 or 4 days after injection of 1 ml of thioglycollate broth. Bone marrow-derived Mφs (BMDMs) were generated by the culture of bone marrow precursors in medium containing macrophage colony-stimulating factor (M-CSF) and total RNA isolated from adherent cells cultured for 6 days. Total RNA (5 µg) was denatured and separated on a 1% formaldehyde–agarose gel prior to capillary transfer to a nylon membrane. Blots were hybridized with [α- 32 P]dATP-labelled DNA probes for β-actin (a 2-kb fragment of human β-actin; Clontech) or hSR-A III transgene (a 1·2-kb fragment of type III hSR-A coding sequence generated by digestion of expression plasmid pcDNA 3 with Hin dIII and Xba I) and exposed to film. (b) Northern blot of adult tissues. Total tissue RNA (10 µg) prepared using organs collected from transgenic line III.4 (Tg + ) mice and non-transgenic (Tg – ) littermates, and from human monocyte-derived Mφ total RNA (10 µg), was denatured and separated on a 1% formaldehyde–agarose gel prior to transfer and hybridization as described in (a). The different sizes of SR-A hybridizing bands in the human and transgenic RNA samples are caused by the absence of 3′ untranslated sequences in the CD68 SR-A type III transgene. (c) Reverse transcription–polymerase chain reaction (RT–PCR) of adult tissues. Total tissue RNA from organs collected from transgenic line III.4 (Tg + ) mice was reverse transcribed in the presence (+) or absence (–) of Moloney murine leukemia virus reverse transcriptase. cDNA synthesized from 100 ng of total RNA served as a template for subsequent PCR using primers specific for hypoxanthine phosphoribosyl transferase (HPRT), macrosialin or the hSR-A III transgene. To enable a comparison of the relative levels of RNA for macrosialin and the hSR-A III transgene, PCR was performed with both oligonucleotide pairs in the same reaction tube. Primers for HPRT and macrosialin do not generate a detectable reaction product from contaminating genomic DNA (gDNA) under the conditions used, while the product corresponding to the amplification of the hSR-A III transgene genomic DNA is 83 bp larger owing to the presence of the first intron (IVS-1) sequence, which is spliced out during the processing of the primary RNA transcript. Reaction products were separated on a 1·2% agarose gel and visualized by ethidium bromide staining.
    Figure Legend Snippet: Analysis of human class A scavenger receptor (hSR-A) III transgene RNA expression. (a) Northern blot of isolated macrophage (Mφ) populations. Total RNA was prepared from peritoneal cells isolated from transgenic line III.11 (Tg + ) mice and non-transgenic (Tg – ) littermates, either 1 or 4 days after injection of 1 ml of thioglycollate broth. Bone marrow-derived Mφs (BMDMs) were generated by the culture of bone marrow precursors in medium containing macrophage colony-stimulating factor (M-CSF) and total RNA isolated from adherent cells cultured for 6 days. Total RNA (5 µg) was denatured and separated on a 1% formaldehyde–agarose gel prior to capillary transfer to a nylon membrane. Blots were hybridized with [α- 32 P]dATP-labelled DNA probes for β-actin (a 2-kb fragment of human β-actin; Clontech) or hSR-A III transgene (a 1·2-kb fragment of type III hSR-A coding sequence generated by digestion of expression plasmid pcDNA 3 with Hin dIII and Xba I) and exposed to film. (b) Northern blot of adult tissues. Total tissue RNA (10 µg) prepared using organs collected from transgenic line III.4 (Tg + ) mice and non-transgenic (Tg – ) littermates, and from human monocyte-derived Mφ total RNA (10 µg), was denatured and separated on a 1% formaldehyde–agarose gel prior to transfer and hybridization as described in (a). The different sizes of SR-A hybridizing bands in the human and transgenic RNA samples are caused by the absence of 3′ untranslated sequences in the CD68 SR-A type III transgene. (c) Reverse transcription–polymerase chain reaction (RT–PCR) of adult tissues. Total tissue RNA from organs collected from transgenic line III.4 (Tg + ) mice was reverse transcribed in the presence (+) or absence (–) of Moloney murine leukemia virus reverse transcriptase. cDNA synthesized from 100 ng of total RNA served as a template for subsequent PCR using primers specific for hypoxanthine phosphoribosyl transferase (HPRT), macrosialin or the hSR-A III transgene. To enable a comparison of the relative levels of RNA for macrosialin and the hSR-A III transgene, PCR was performed with both oligonucleotide pairs in the same reaction tube. Primers for HPRT and macrosialin do not generate a detectable reaction product from contaminating genomic DNA (gDNA) under the conditions used, while the product corresponding to the amplification of the hSR-A III transgene genomic DNA is 83 bp larger owing to the presence of the first intron (IVS-1) sequence, which is spliced out during the processing of the primary RNA transcript. Reaction products were separated on a 1·2% agarose gel and visualized by ethidium bromide staining.

    Techniques Used: RNA Expression, Northern Blot, Isolation, Transgenic Assay, Mouse Assay, Injection, Derivative Assay, Generated, Cell Culture, Agarose Gel Electrophoresis, Sequencing, Expressing, Plasmid Preparation, Hybridization, Reverse Transcription Polymerase Chain Reaction, Synthesized, Polymerase Chain Reaction, Amplification, Staining

    Related Articles

    Amplification:

    Article Title: Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2
    Article Snippet: .. Reverse transcription–polymerase chain reaction (RT)–PCR Total RNA was extracted from the cells using MRC Trizol reagent (Cincinnati, OH, USA). cDNA was prepared from 1 μ g of total RNA using oligo(dT) primer and Moloney murine leukaemia virus reverse transcriptase (Gibco BRL), and amplified by PCR with Taq DNA polymerase. ..

    Synthesized:

    Article Title: Proanthocyanidin oxidation of Arabidopsis seeds is altered in mutant of the high-affinity nitrate transporter NRT2.7
    Article Snippet: .. First-strand cDNAs were synthesized from 1 μg RNA using Moloney murine leukaemia virus reverse transcriptase (Thermo Scientific) and oligo(dT)15 primers (Thermo Scientific). .. The absence of DNA contamination was verified by PCR using specific primers spanning an intron in Nii (At2g15620): forward 5′-TGCTGATGACGTTCTTCCACTCTGC-3′; reverse 5′-CTG AGG GTT GACTCCGAAATA GTCTC-3′.

    Sequencing:

    Article Title: Role of PECAM-1 in radiation-induced liver inflammation
    Article Snippet: .. Briefly, cDNA was generated by reverse transcription of 1 μg of total RNA using 100 nM of dNTPs, 50 pM of primer oligo dT15, 200 U of moloney murine leukaemia virus reverse transcriptase (M-MLV RT), 16 U of protector RNase inhibitor in RT-buffer and 2.5 μl of 0.1 M DTT; real-time PCR was performed with a StepOnePlus– sequence detection system (Applied Biosystems, Darmstadt, Germany) as described previously with the primers shown in . .. Fold change expression was calculated using threshold cycle (Ct) values.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2
    Article Snippet: .. Reverse transcription–polymerase chain reaction (RT)–PCR Total RNA was extracted from the cells using MRC Trizol reagent (Cincinnati, OH, USA). cDNA was prepared from 1 μ g of total RNA using oligo(dT) primer and Moloney murine leukaemia virus reverse transcriptase (Gibco BRL), and amplified by PCR with Taq DNA polymerase. ..

    Article Title: The use of human CD68 transcriptional regulatory sequences to direct high-level expression of class A scavenger receptor in macrophages in vitro and in vivo
    Article Snippet: .. For reverse transcription–PCR (RT–PCR) analyses, total RNA was reverse transcribed using Moloney murine leukaemia virus reverse transcriptase and an Oligo-dT primer (Life Technologies), with reactions set up in the presence and absence of reverse transcriptase to control for genomic DNA contamination. .. The cDNA obtained served as a template for PCR using the following oligonucleotide pairs: HPRT: 5′-GCTACCTGCTGGATTACAT-3′ and 5′-CCAGTTTCACTAATGACACAA-3′; macrosialin: 5′-TCCTTCACGATGACACCTACAG-3′ and 5′-GGACCAGGCCAATGATGAGAG-3′; hSR-A III transgene: 5′-GGGTGAGGCGGTTCAGCC-3′ and 5′-TTTCCTCTTCGCTGTCATTTC-3′.

    Real-time Polymerase Chain Reaction:

    Article Title: Role of PECAM-1 in radiation-induced liver inflammation
    Article Snippet: .. Briefly, cDNA was generated by reverse transcription of 1 μg of total RNA using 100 nM of dNTPs, 50 pM of primer oligo dT15, 200 U of moloney murine leukaemia virus reverse transcriptase (M-MLV RT), 16 U of protector RNase inhibitor in RT-buffer and 2.5 μl of 0.1 M DTT; real-time PCR was performed with a StepOnePlus– sequence detection system (Applied Biosystems, Darmstadt, Germany) as described previously with the primers shown in . .. Fold change expression was calculated using threshold cycle (Ct) values.

    Polymerase Chain Reaction:

    Article Title: Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2
    Article Snippet: .. Reverse transcription–polymerase chain reaction (RT)–PCR Total RNA was extracted from the cells using MRC Trizol reagent (Cincinnati, OH, USA). cDNA was prepared from 1 μ g of total RNA using oligo(dT) primer and Moloney murine leukaemia virus reverse transcriptase (Gibco BRL), and amplified by PCR with Taq DNA polymerase. ..

    Article Title: Contrasting molecular composition and channel properties of AMPA receptors on chick auditory and brainstem motor neurons
    Article Snippet: .. For reverse transcription, this RNA was used in a reverse transcription reaction with Superscript II, a modified Moloney murine leukaemia virus reverse transcriptase (Gibco BRL), and an oligo-dT primer to produce cDNA for use as templates in PCR. .. PCR of a homologous region of GluR1-4 was performed using PCR Supermix (Gibco BRL) and the pan-AMPA degenerate primers listed below, according to the manufacturer's protocol.

    Generated:

    Article Title: Role of PECAM-1 in radiation-induced liver inflammation
    Article Snippet: .. Briefly, cDNA was generated by reverse transcription of 1 μg of total RNA using 100 nM of dNTPs, 50 pM of primer oligo dT15, 200 U of moloney murine leukaemia virus reverse transcriptase (M-MLV RT), 16 U of protector RNase inhibitor in RT-buffer and 2.5 μl of 0.1 M DTT; real-time PCR was performed with a StepOnePlus– sequence detection system (Applied Biosystems, Darmstadt, Germany) as described previously with the primers shown in . .. Fold change expression was calculated using threshold cycle (Ct) values.

    Modification:

    Article Title: Contrasting molecular composition and channel properties of AMPA receptors on chick auditory and brainstem motor neurons
    Article Snippet: .. For reverse transcription, this RNA was used in a reverse transcription reaction with Superscript II, a modified Moloney murine leukaemia virus reverse transcriptase (Gibco BRL), and an oligo-dT primer to produce cDNA for use as templates in PCR. .. PCR of a homologous region of GluR1-4 was performed using PCR Supermix (Gibco BRL) and the pan-AMPA degenerate primers listed below, according to the manufacturer's protocol.

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