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primer antisense mix  (New England Biolabs)


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    Structured Review

    New England Biolabs primer antisense mix
    Primer Antisense Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 1645 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/antisense+primer/Random+Primer+Mix/pm40062677-82-22-38
    Average 97 stars, based on 1645 article reviews
    primer antisense mix - by Bioz Stars, 2026-10
    97/100 stars

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    Related Articles

    cDNA Synthesis:

    Article Title: Nuclear envelope budding enables export of large transcripts in muscle cells.
    Article Snippet: RNA was eluted in 50 μl of nuclease-free water and the concentration was determined using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific). .. First-strand cDNA synthesis was performed on all RNA samples using the LunaScript® RT Master Mix Kit (NEB #E3025L) combined with the Random Primer Mix (NEB #S1330S) according to the manufacturer’s included protocol. ..

    Article Title: Essential role of cytochrome bc 1 in Pseudomonas aeruginosa cell physiology and virulence
    Article Snippet: .. Supernatant total RNA was extracted using TRIzol LS (Thermo Fisher Scientific, Waltham, MA) and quantified on a Qubit 3.0 fluorometer using the AccuBlue Broad Range RNA Quantitation Kit (Biotium, Fremont, CA), followed by cDNA synthesis with the ProtoScript II First Strand cDNA Synthesis Kit using the Random Primer Mix (New England Biolabs, Ipswich, MA). qPCR was subsequently performed using the primer pairs listed in , designed using Primer 3 software ( ; ; ). .. P. aeruginosa rpoD was utilized as a housekeeping gene due to its vital role in promoter identification to facilitate constitutive gene expression ( ; ). cDNA (100 ng) amplification and quantification were performed on a Bio-Rad CFX Connect Real-Time PCR Detection System using the iTaq Universal SYBR Green Supermix kit (Bio-Rad, Hercules, CA). mRNA expression was determined via a 2-ΔΔCT method comparing rpoD , exoT , exoU , and exoY expression in the presence of P. aeruginosa PA14 or PA14 Δ bc 1 infection via cycle threshold analysis ( ).

    Construct:

    Article Title: Metabolic and behavioral effects of neurofibromin result from differential recruitment of MAPK and mTOR signaling.
    Article Snippet: RNA was isolated with the Qiagen RNeasy Lipid Tissue Mini Kit. .. Complimentary DNA library was constructed using LunaScript RT Master Mix (New England Biolabs, catalog #E3025L) with the Random Primer Mix (New England Biolabs, catalog #S1330S). .. Nf1 gene expression was quantified in fly head samples using Luna Universal qPCR Master Mix (New England Biolabs, catalog #M3003L).

    Article Title: Metabolic and behavioral effects of neurofibromin result from differential recruitment of MAPK and mTOR signaling
    Article Snippet: RNA was isolated with the Qiagen RNeasy Lipid Tissue Mini Kit. .. Complimentary DNA library was constructed using LunaScript RT Master Mix (New England Biolabs, catalog #E3025L) with the Random Primer Mix (New England Biolabs, catalog #S1330S). .. Nf1 gene expression was quantified in fly head samples using Luna Universal qPCR Master Mix (New England Biolabs, catalog #M3003L).

    Quantitative RT-PCR:

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system.
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix LoROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix Lo-ROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Reverse Transcription:

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system.
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix LoROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix Lo-ROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Article Title: Comparative Analysis of Prenatal Stress Models: Placental and Neurodevelopmental Outcomes in Mice
    Article Snippet: .. E14 and E18 placenta and ventral forebrain from one male and one female per litter were randomly selected for RNA isolation (Trizol) and quantified and verified by Spectrophotometer (Nanodrop). cDNA was synthesized (New England Biolabs AMV Reverse Transcriptase; NEB M0227L, Random Primer Mix; NEB S1330S). ..

    Random Hexamer:

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system.
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix LoROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system
    Article Snippet: .. The cDNA for the RT-qPCR was prepared by reverse transcription of the extracted total RNA and a mixture of random hexamer primers and oligoT primers (random primer mix, New England Biolabs, Ipswich, MA, USA, #S1330S) using RevertAid Reverse Transcriptase (Thermo Scientific, Vilnius, Lithuania, #EP0442) following the manufacturerʼs instructions. .. The amplification reaction for each gene was performed in a 10 μl volume containing 1 x SG Master Mix (Xceed qPCR SG 2x Mix Lo-ROX, IAB, Prague, Czech Republic, #LPCR10501XL), 0.5 μM forward/reverse primer, and 3 μl of the sample containing the cDNA template.

    Quantitation Assay:

    Article Title: Essential role of cytochrome bc 1 in Pseudomonas aeruginosa cell physiology and virulence
    Article Snippet: .. Supernatant total RNA was extracted using TRIzol LS (Thermo Fisher Scientific, Waltham, MA) and quantified on a Qubit 3.0 fluorometer using the AccuBlue Broad Range RNA Quantitation Kit (Biotium, Fremont, CA), followed by cDNA synthesis with the ProtoScript II First Strand cDNA Synthesis Kit using the Random Primer Mix (New England Biolabs, Ipswich, MA). qPCR was subsequently performed using the primer pairs listed in , designed using Primer 3 software ( ; ; ). .. P. aeruginosa rpoD was utilized as a housekeeping gene due to its vital role in promoter identification to facilitate constitutive gene expression ( ; ). cDNA (100 ng) amplification and quantification were performed on a Bio-Rad CFX Connect Real-Time PCR Detection System using the iTaq Universal SYBR Green Supermix kit (Bio-Rad, Hercules, CA). mRNA expression was determined via a 2-ΔΔCT method comparing rpoD , exoT , exoU , and exoY expression in the presence of P. aeruginosa PA14 or PA14 Δ bc 1 infection via cycle threshold analysis ( ).

    Real-time Polymerase Chain Reaction:

    Article Title: Essential role of cytochrome bc 1 in Pseudomonas aeruginosa cell physiology and virulence
    Article Snippet: .. Supernatant total RNA was extracted using TRIzol LS (Thermo Fisher Scientific, Waltham, MA) and quantified on a Qubit 3.0 fluorometer using the AccuBlue Broad Range RNA Quantitation Kit (Biotium, Fremont, CA), followed by cDNA synthesis with the ProtoScript II First Strand cDNA Synthesis Kit using the Random Primer Mix (New England Biolabs, Ipswich, MA). qPCR was subsequently performed using the primer pairs listed in , designed using Primer 3 software ( ; ; ). .. P. aeruginosa rpoD was utilized as a housekeeping gene due to its vital role in promoter identification to facilitate constitutive gene expression ( ; ). cDNA (100 ng) amplification and quantification were performed on a Bio-Rad CFX Connect Real-Time PCR Detection System using the iTaq Universal SYBR Green Supermix kit (Bio-Rad, Hercules, CA). mRNA expression was determined via a 2-ΔΔCT method comparing rpoD , exoT , exoU , and exoY expression in the presence of P. aeruginosa PA14 or PA14 Δ bc 1 infection via cycle threshold analysis ( ).

    Software:

    Article Title: Essential role of cytochrome bc 1 in Pseudomonas aeruginosa cell physiology and virulence
    Article Snippet: .. Supernatant total RNA was extracted using TRIzol LS (Thermo Fisher Scientific, Waltham, MA) and quantified on a Qubit 3.0 fluorometer using the AccuBlue Broad Range RNA Quantitation Kit (Biotium, Fremont, CA), followed by cDNA synthesis with the ProtoScript II First Strand cDNA Synthesis Kit using the Random Primer Mix (New England Biolabs, Ipswich, MA). qPCR was subsequently performed using the primer pairs listed in , designed using Primer 3 software ( ; ; ). .. P. aeruginosa rpoD was utilized as a housekeeping gene due to its vital role in promoter identification to facilitate constitutive gene expression ( ; ). cDNA (100 ng) amplification and quantification were performed on a Bio-Rad CFX Connect Real-Time PCR Detection System using the iTaq Universal SYBR Green Supermix kit (Bio-Rad, Hercules, CA). mRNA expression was determined via a 2-ΔΔCT method comparing rpoD , exoT , exoU , and exoY expression in the presence of P. aeruginosa PA14 or PA14 Δ bc 1 infection via cycle threshold analysis ( ).

    Isolation:

    Article Title: Comparative Analysis of Prenatal Stress Models: Placental and Neurodevelopmental Outcomes in Mice
    Article Snippet: .. E14 and E18 placenta and ventral forebrain from one male and one female per litter were randomly selected for RNA isolation (Trizol) and quantified and verified by Spectrophotometer (Nanodrop). cDNA was synthesized (New England Biolabs AMV Reverse Transcriptase; NEB M0227L, Random Primer Mix; NEB S1330S). ..

    Spectrophotometry:

    Article Title: Comparative Analysis of Prenatal Stress Models: Placental and Neurodevelopmental Outcomes in Mice
    Article Snippet: .. E14 and E18 placenta and ventral forebrain from one male and one female per litter were randomly selected for RNA isolation (Trizol) and quantified and verified by Spectrophotometer (Nanodrop). cDNA was synthesized (New England Biolabs AMV Reverse Transcriptase; NEB M0227L, Random Primer Mix; NEB S1330S). ..

    Synthesized:

    Article Title: Comparative Analysis of Prenatal Stress Models: Placental and Neurodevelopmental Outcomes in Mice
    Article Snippet: .. E14 and E18 placenta and ventral forebrain from one male and one female per litter were randomly selected for RNA isolation (Trizol) and quantified and verified by Spectrophotometer (Nanodrop). cDNA was synthesized (New England Biolabs AMV Reverse Transcriptase; NEB M0227L, Random Primer Mix; NEB S1330S). ..



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    Image Search Results


    FASN constructions used in this study. A. The delimitation of each domain is indicated by numbering the amino-acids. Each construction encodes a Flag-peptide located at the N-terminus. The domain boundaries shown are based on the UniProt primary sequence annotation (fatty acid synthase access number P49327 ). B. Domain boundaries shown are based on available experimental structures (PDB IDs: 3HHD , 8G7X , 4PIV , 4w82 and 7mhd ) ACP, Acyl Carrier Protein; DH, DeHydrogenase; ER, Enoyl-Reductase; KR, Keto-acyl Reductase; KS, Keto-acyl Synthase; MAT, Malonyl/Acetyl Transferase; TE, ThioEsterase.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Mammalian fatty acid synthase and O -GlcNAc transferase preferentially interact via their respective N -terminal regions

    doi: 10.1016/j.bbrep.2025.102427

    Figure Lengend Snippet: FASN constructions used in this study. A. The delimitation of each domain is indicated by numbering the amino-acids. Each construction encodes a Flag-peptide located at the N-terminus. The domain boundaries shown are based on the UniProt primary sequence annotation (fatty acid synthase access number P49327 ). B. Domain boundaries shown are based on available experimental structures (PDB IDs: 3HHD , 8G7X , 4PIV , 4w82 and 7mhd ) ACP, Acyl Carrier Protein; DH, DeHydrogenase; ER, Enoyl-Reductase; KR, Keto-acyl Reductase; KS, Keto-acyl Synthase; MAT, Malonyl/Acetyl Transferase; TE, ThioEsterase.

    Article Snippet: FASN gene was cloned into the pCMV10-3xFlag expression vector thanks to the Hin d III - FASN sense primer and FASN - Xba I antisense primer (Eurogentec) ( ).

    Techniques: Sequencing

    FASN and OGT interact via their respective N-terminal part. A and B. Hep3B cells were transfected with an empty vector (A) or with the different 3xFlag-FASN deletion mutants (B to G) in combination with OGT-HA overexpression. Protein samples were incubated with an anti-Flag antibody for co-immunoprecipitation with OGT (“IP” lanes for “Immuno-Precipitated”) ( panel A ) or with WGA-agarose beads (“P” lanes for “Purified”) ( panel B ). A. Co-immunoprecipitation of the deletion mutants with OGT analyzed by Western blotting. Optical densities of co-IP OGT were measured and normalized with the corresponding IP deletion mutants of FASN (red arrows; n = 3) (arbitrary units). B. O -GlcNAcylation levels of the deletion mutants analyzed by Western blotting. Specificity of WGA was checked by adding 0.5 M GlcNAc (“P + Gn” lane). Optical densities of the “purified” O -GlcNAcylated constructs were measured and normalized with inputs (red arrows; n = 3) (arbitrary units). ∗ p < 0.05; ∗∗ p < 0.01. C. Hep3B cells were transfected with 3xFlag-FASN. Protein samples were incubated with a GST-OGT construct (1–3) or GST only (4). After GST pull-down, interaction of the GST-OGT constructs (red arrows) with 3xFlag-FASN was analyzed by Western blotting. Optical densities of pull-down 3xFlag-FASN were measured and normalized with the corresponding construct. Interaction with GST only was subtracted from the other bands (n = 4). CD, Catalytic Domain; GST, Gluthatione-S Transferase; NLS, Nuclear Localization Signal; TPR, TetratricoPeptide Repeats. Dotted lines indicate where the blots were cut and joined during figure preparation. Molecular mass markers are indicated on the left (kDa).

    Journal: Biochemistry and Biophysics Reports

    Article Title: Mammalian fatty acid synthase and O -GlcNAc transferase preferentially interact via their respective N -terminal regions

    doi: 10.1016/j.bbrep.2025.102427

    Figure Lengend Snippet: FASN and OGT interact via their respective N-terminal part. A and B. Hep3B cells were transfected with an empty vector (A) or with the different 3xFlag-FASN deletion mutants (B to G) in combination with OGT-HA overexpression. Protein samples were incubated with an anti-Flag antibody for co-immunoprecipitation with OGT (“IP” lanes for “Immuno-Precipitated”) ( panel A ) or with WGA-agarose beads (“P” lanes for “Purified”) ( panel B ). A. Co-immunoprecipitation of the deletion mutants with OGT analyzed by Western blotting. Optical densities of co-IP OGT were measured and normalized with the corresponding IP deletion mutants of FASN (red arrows; n = 3) (arbitrary units). B. O -GlcNAcylation levels of the deletion mutants analyzed by Western blotting. Specificity of WGA was checked by adding 0.5 M GlcNAc (“P + Gn” lane). Optical densities of the “purified” O -GlcNAcylated constructs were measured and normalized with inputs (red arrows; n = 3) (arbitrary units). ∗ p < 0.05; ∗∗ p < 0.01. C. Hep3B cells were transfected with 3xFlag-FASN. Protein samples were incubated with a GST-OGT construct (1–3) or GST only (4). After GST pull-down, interaction of the GST-OGT constructs (red arrows) with 3xFlag-FASN was analyzed by Western blotting. Optical densities of pull-down 3xFlag-FASN were measured and normalized with the corresponding construct. Interaction with GST only was subtracted from the other bands (n = 4). CD, Catalytic Domain; GST, Gluthatione-S Transferase; NLS, Nuclear Localization Signal; TPR, TetratricoPeptide Repeats. Dotted lines indicate where the blots were cut and joined during figure preparation. Molecular mass markers are indicated on the left (kDa).

    Article Snippet: FASN gene was cloned into the pCMV10-3xFlag expression vector thanks to the Hin d III - FASN sense primer and FASN - Xba I antisense primer (Eurogentec) ( ).

    Techniques: Transfection, Plasmid Preparation, Over Expression, Incubation, Immunoprecipitation, Purification, Western Blot, Co-Immunoprecipitation Assay, Construct

    FASN and OGT interaction modeling. A. Modeling of FASN dimer and two OGT monomers; full sequences of the FASN dimer and of the OGT monomer were modeled independently with AlphaFold (see Materials and methods), assembled according to co-prediction performed in 3C and rendered in cartoon representation, FASN dimer in magenta and green (2511 residues each) and OGT monomers in cyan (1046 residues each). B. Same as A, but with FASN colored according to the domains depicted in and the four O -GlcNAcylated sites previously identified and reported, T315, S806, T980 and S1534, highlighted by red spheres . C. Model of FASN N -ter (bottom domain) and OGT N -ter (top domain, the TPR helices are clearly visible); the FASN sequence was trimmed to 1–969 and the OGT sequence to 1–286; the structure was predicted using AlphaFold3 and rendered in cartoon representation; residues are colored according to the pLDDT confidence score. D . Predicted Aligned Error matrix of the complex in C , ordered as OGT first and then FASN.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Mammalian fatty acid synthase and O -GlcNAc transferase preferentially interact via their respective N -terminal regions

    doi: 10.1016/j.bbrep.2025.102427

    Figure Lengend Snippet: FASN and OGT interaction modeling. A. Modeling of FASN dimer and two OGT monomers; full sequences of the FASN dimer and of the OGT monomer were modeled independently with AlphaFold (see Materials and methods), assembled according to co-prediction performed in 3C and rendered in cartoon representation, FASN dimer in magenta and green (2511 residues each) and OGT monomers in cyan (1046 residues each). B. Same as A, but with FASN colored according to the domains depicted in and the four O -GlcNAcylated sites previously identified and reported, T315, S806, T980 and S1534, highlighted by red spheres . C. Model of FASN N -ter (bottom domain) and OGT N -ter (top domain, the TPR helices are clearly visible); the FASN sequence was trimmed to 1–969 and the OGT sequence to 1–286; the structure was predicted using AlphaFold3 and rendered in cartoon representation; residues are colored according to the pLDDT confidence score. D . Predicted Aligned Error matrix of the complex in C , ordered as OGT first and then FASN.

    Article Snippet: FASN gene was cloned into the pCMV10-3xFlag expression vector thanks to the Hin d III - FASN sense primer and FASN - Xba I antisense primer (Eurogentec) ( ).

    Techniques: Sequencing