rapid dna ligation kit  (Thermo Fisher)


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    Name:
    Rapid DNA Ligation Kit
    Description:
    Thermo Scientific Rapid DNA Ligation Kit enables fast sticky end or blunt end DNA ligation in only 5 minutes at room temperature The kit contains T4 DNA ligase and a specially formulated 5X rapid ligation buffer optimized for fast and efficient DNA ligation Fast ligation efficiency is equal to that obtained with T4 DNA ligase in a standard 1 hour ligation The ligation reaction mixture can be used directly for bacterial transformation with the TransformAid Bacterial Transformation Kit or with other conventional transformation procedures Highlights• Fast ligation of either sticky or blunt end DNA in only 5 minutes• Convenient reaction mixture can be used directly for bacterial transformationApplications• Routine cloning experiments• Blunt end cloning• Library construction• TA cloningIncludes• T4 DNA Ligase• 5X Rapid Ligation Buffer• Water nuclease free• Detailed ProtocolNotePrior to electroporation it is necessary to inactivate T4 DNA ligase by chloroform extraction or spin column purification e g with Thermo Scientific GeneJET PCR Purification Kit K0701 Related ProductsRapid DNA Ligation Kit
    Catalog Number:
    K1422
    Price:
    None
    Category:
    Proteins Enzymes Peptides
    Applications:
    Cloning|PCR Cloning|Restriction Enzyme Cloning
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    Structured Review

    Thermo Fisher rapid dna ligation kit
    Thermo Scientific Rapid DNA Ligation Kit enables fast sticky end or blunt end DNA ligation in only 5 minutes at room temperature The kit contains T4 DNA ligase and a specially formulated 5X rapid ligation buffer optimized for fast and efficient DNA ligation Fast ligation efficiency is equal to that obtained with T4 DNA ligase in a standard 1 hour ligation The ligation reaction mixture can be used directly for bacterial transformation with the TransformAid Bacterial Transformation Kit or with other conventional transformation procedures Highlights• Fast ligation of either sticky or blunt end DNA in only 5 minutes• Convenient reaction mixture can be used directly for bacterial transformationApplications• Routine cloning experiments• Blunt end cloning• Library construction• TA cloningIncludes• T4 DNA Ligase• 5X Rapid Ligation Buffer• Water nuclease free• Detailed ProtocolNotePrior to electroporation it is necessary to inactivate T4 DNA ligase by chloroform extraction or spin column purification e g with Thermo Scientific GeneJET PCR Purification Kit K0701 Related ProductsRapid DNA Ligation Kit
    https://www.bioz.com/result/rapid dna ligation kit/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rapid dna ligation kit - by Bioz Stars, 2021-05
    98/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: 5-Lipoxygenase and Leukotriene B4 Receptor Are Expressed in Human Pancreatic Cancers But Not in Pancreatic Ducts in Normal Tissue
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody, and Luminol reagents were from New England BioLabs (Beverly, MA). .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, the diaminobenzidine reagent set, and the Histo-Mark-Red AP-system were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Article Title: FlgM as a Secretion Moiety for the Development of an Inducible Type III Secretion System
    Article Snippet: .. Rapid DNA ligation kit, Pfu DNA polymerase and 10× MgSO4 -PCR buffer were obtained from Fermentas. .. Tris-Glycine gels were purchased from Invitrogen and Biorad.

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: Subsequently, the EF1α promoter was amplified from the pEF1α-IRES vector (Clontech) by PCR using primers containing NruI/NheI sites. .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Differential Expression of Thromboxane Synthase in Prostate Carcinoma
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody and Luminol reagents were from Amersham. .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, and the diaminobenzidine reagent set were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Clone Assay:

    Article Title: 5-Lipoxygenase and Leukotriene B4 Receptor Are Expressed in Human Pancreatic Cancers But Not in Pancreatic Ducts in Normal Tissue
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody, and Luminol reagents were from New England BioLabs (Beverly, MA). .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, the diaminobenzidine reagent set, and the Histo-Mark-Red AP-system were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Article Title: Differential Expression of Thromboxane Synthase in Prostate Carcinoma
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody and Luminol reagents were from Amersham. .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, and the diaminobenzidine reagent set were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Plasmid Preparation:

    Article Title: 5-Lipoxygenase and Leukotriene B4 Receptor Are Expressed in Human Pancreatic Cancers But Not in Pancreatic Ducts in Normal Tissue
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody, and Luminol reagents were from New England BioLabs (Beverly, MA). .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, the diaminobenzidine reagent set, and the Histo-Mark-Red AP-system were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Article Title: KIF14 controls ciliogenesis via regulation of Aurora A and is important for Hedgehog signaling
    Article Snippet: DNA constructs and stable cell linesTo generate destination vector with N-terminal MYC-BirA* tag, MYC-BirA* was amplified by PCR from pcDNA3.1 MYC-BirA* (43920; Addgene) using forward primer NO°1 and reverse primer NO°2 ( ) to create AflII and BstBI site–containing overhangs and cloned into TOPO entry vector (using TOPO TA Cloning Kit, K450001; ThermoFisher) sites. .. Next, following digest with AflII and BstBI (enzymes from NEB) the MYC-BirA* fragment was ligated (K1422; ThermoFisher) into AflII and BstBI sites of pgLAP2 vector (19703; Addgene). .. MYC-BirA*-CEP164 construct was generated from pDONOR-CEP164 ( ) and MYC-BirA* destination vector described above by LR Gateway cloning reaction (ThermoFisher), and subsequently validated for correct expression and localization by Western blot and immunofluorescence microscopy analyses following transfection of HEK293T cells.

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: Subsequently, the EF1α promoter was amplified from the pEF1α-IRES vector (Clontech) by PCR using primers containing NruI/NheI sites. .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Differential Expression of Thromboxane Synthase in Prostate Carcinoma
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody and Luminol reagents were from Amersham. .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, and the diaminobenzidine reagent set were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    DNA Ligation:

    Article Title: 5-Lipoxygenase and Leukotriene B4 Receptor Are Expressed in Human Pancreatic Cancers But Not in Pancreatic Ducts in Normal Tissue
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody, and Luminol reagents were from New England BioLabs (Beverly, MA). .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, the diaminobenzidine reagent set, and the Histo-Mark-Red AP-system were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Article Title: Transposable Elements as Stress Adaptive Capacitors Induce Genomic Instability in Fungal Pathogen Magnaporthe oryzae
    Article Snippet: All oligonucleotides based on DNA sequences of transposons and retrotransposons were synthesized by Metabion International AG (Germany). .. GeneRuler DNA markers and Fast-Link DNA ligation kit were purchased from Fermentas and Roche Diagnostics (Mannheim, Germany), respectively. .. Plasmid DNA was isolated from E. coli by the alkaline lysis method, employing a QIAprep spin miniprep kit from Qiagen.

    Article Title: FlgM as a Secretion Moiety for the Development of an Inducible Type III Secretion System
    Article Snippet: .. Rapid DNA ligation kit, Pfu DNA polymerase and 10× MgSO4 -PCR buffer were obtained from Fermentas. .. Tris-Glycine gels were purchased from Invitrogen and Biorad.

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: Subsequently, the EF1α promoter was amplified from the pEF1α-IRES vector (Clontech) by PCR using primers containing NruI/NheI sites. .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Article Title: Differential Expression of Thromboxane Synthase in Prostate Carcinoma
    Article Snippet: The horseradish peroxidase-conjugated secondary antibody and Luminol reagents were from Amersham. .. The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA). .. The normal goat serum, the streptavidin-peroxidase, the streptavidin-phosphatase, and the diaminobenzidine reagent set were purchased from Kirkegaard & Perry Laboratories (Gaithersburg, MD).

    Article Title: A combination of transposable elements and magnetic cell sorting provides a very efficient transgenesis system for chicken primary erythroid progenitors
    Article Snippet: This digestion was performed using the Fermentas 10× FastDigest buffer, in a final volume of 20 μl. .. Considering that a 4 bp recognition site will occur roughly every 256 bp, and therefore that 200 ng of Tai I-digested gDNA correspond to 1.28 pmol DNA, 200 ng of digested DNA were ligated to a 3× molar excess of the annealed splinkerette adaptor (Splinklong-ACGT+Splinkshort-hairpin) with the T4 DNA ligase (4 U), for 1 hour at 22°C, using the Fermentas Rapid DNA Ligation kit. .. In order to improve the ligation yield, digested DNA was heated for 5 min at 60°C before being ligated.

    Amplification:

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: Subsequently, the EF1α promoter was amplified from the pEF1α-IRES vector (Clontech) by PCR using primers containing NruI/NheI sites. .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

    Purification:

    Article Title: Generating aldehyde-tagged antibodies with high titers and high formylglycine yields by supplementing culture media with copper(II)
    Article Snippet: Subsequently, the EF1α promoter was amplified from the pEF1α-IRES vector (Clontech) by PCR using primers containing NruI/NheI sites. .. Both the FGE vector and the EF1α PCR product were digested with NruI and NheI and ligated with the Rapid DNA Ligation Kit (Thermo/Life Technologies) to generate WT pEF-FGE-hygro, which was amplified in XL-10 gold cells (Agilent) and purified according to the manufacturer’s instructions. .. Quikchange Mutagenesis (Agilent) was used to mutate the furin cleavage site (HRYSRE to HRYSGE) and/or add the KDEL retention signal to the C-terminus of FGE to generate Δfurin, +KDEL, and Δfurin + KDEL versions of FGE.

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    Thermo Fisher fast link dna ligation kit
    Genetic variability among M. oryzae isolates. <t>DNA</t> profiles were obtained for M. oryzae isolates using outward primers PyR1 (upper) and Pot2L2 (lower) derived from LTR-retrotransposon Pyret and DNA transposon Pot2 respectively. Details of PyR1 and Pot2L2 primers are provided in table 1 . Lanes 1 to 9 represents M. oryzae isolates from similar agro-climatic region of India ( Table 3 ). Lane M represents Fermentas <t>GeneRuler</t> 100 bp Plus DNA Ladder. As compared to Pot2, Pyret generated high intra-regional genetic variations among M. oryzae isolate.
    Fast Link Dna Ligation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fast link dna ligation kit/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fast link dna ligation kit - by Bioz Stars, 2021-05
    98/100 stars
      Buy from Supplier

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    Genetic variability among M. oryzae isolates. DNA profiles were obtained for M. oryzae isolates using outward primers PyR1 (upper) and Pot2L2 (lower) derived from LTR-retrotransposon Pyret and DNA transposon Pot2 respectively. Details of PyR1 and Pot2L2 primers are provided in table 1 . Lanes 1 to 9 represents M. oryzae isolates from similar agro-climatic region of India ( Table 3 ). Lane M represents Fermentas GeneRuler 100 bp Plus DNA Ladder. As compared to Pot2, Pyret generated high intra-regional genetic variations among M. oryzae isolate.

    Journal: PLoS ONE

    Article Title: Transposable Elements as Stress Adaptive Capacitors Induce Genomic Instability in Fungal Pathogen Magnaporthe oryzae

    doi: 10.1371/journal.pone.0094415

    Figure Lengend Snippet: Genetic variability among M. oryzae isolates. DNA profiles were obtained for M. oryzae isolates using outward primers PyR1 (upper) and Pot2L2 (lower) derived from LTR-retrotransposon Pyret and DNA transposon Pot2 respectively. Details of PyR1 and Pot2L2 primers are provided in table 1 . Lanes 1 to 9 represents M. oryzae isolates from similar agro-climatic region of India ( Table 3 ). Lane M represents Fermentas GeneRuler 100 bp Plus DNA Ladder. As compared to Pot2, Pyret generated high intra-regional genetic variations among M. oryzae isolate.

    Article Snippet: GeneRuler DNA markers and Fast-Link DNA ligation kit were purchased from Fermentas and Roche Diagnostics (Mannheim, Germany), respectively.

    Techniques: Derivative Assay, Generated

    KIF14 depletion deregulates AURA. (A) Representative images of IF detection of pAURA (green), Ac-tub (red), CAP350 (yellow), and DNA (blue) in serum-starved hTERT RPE-1 cells; scale bar, 5 µm. (B) Quantification of pAURA fluorescent intensity (normalized to Hoechst signal intensity) in serum-starved whole cells. (C) Detection of a BB-specific pool of pAURA (green) in nonstarved cells using high-resolution microscopy (CAP350, blue; Ac-tub, red); scale bar, 0,4 µm. (D) Quantification of pAURA signal intensity on BB (normalized to CAP350 signal intensity). (E–L) Effect of MYC-AURA overexpression on 24 h serum-starved hTERT RPE-1 cells. (E) Representative images of IF detection of MYC-tagged (green), Ac-tub + primary cilia (red; CAP350, yellow; DNA, blue) in mock- or MYC-AURA–transfected cells; scale bar, 10 µm. (F) Graph quantifying statistically significant decrease of ciliated cells percentage after MYC-AURA overexpression. (G and H) Examination of FBF1 (green) localization and decreased intensity after MYC-AURA overexpression. Representative images (CAP350 in red, MYC in yellow) are shown in G (scale bar, 2 µm) and intensity quantification (normalized to CAP350) in H. (I and J) Examination of SCLT1 (green) localization and decreased intensity after MYC-AURA overexpression. Representative images (CAP350 in red, MYC in yellow) are shown in I (scale bar, 1 µm) and intensity quantification (normalized to CAP350) in J. (K and L) Examination of IFT57 (green) localization and decreased intensity at BBs after MYC-AURA overexpression. Representative images (Ac-tub in red, CAP350 in yellow) are shown in K (scale bar, 2 µm) and BB intensity quantification (normalized to CAP350) in L. Asterisks indicate statistical significance determined using an unpaired t test.

    Journal: The Journal of Cell Biology

    Article Title: KIF14 controls ciliogenesis via regulation of Aurora A and is important for Hedgehog signaling

    doi: 10.1083/jcb.201904107

    Figure Lengend Snippet: KIF14 depletion deregulates AURA. (A) Representative images of IF detection of pAURA (green), Ac-tub (red), CAP350 (yellow), and DNA (blue) in serum-starved hTERT RPE-1 cells; scale bar, 5 µm. (B) Quantification of pAURA fluorescent intensity (normalized to Hoechst signal intensity) in serum-starved whole cells. (C) Detection of a BB-specific pool of pAURA (green) in nonstarved cells using high-resolution microscopy (CAP350, blue; Ac-tub, red); scale bar, 0,4 µm. (D) Quantification of pAURA signal intensity on BB (normalized to CAP350 signal intensity). (E–L) Effect of MYC-AURA overexpression on 24 h serum-starved hTERT RPE-1 cells. (E) Representative images of IF detection of MYC-tagged (green), Ac-tub + primary cilia (red; CAP350, yellow; DNA, blue) in mock- or MYC-AURA–transfected cells; scale bar, 10 µm. (F) Graph quantifying statistically significant decrease of ciliated cells percentage after MYC-AURA overexpression. (G and H) Examination of FBF1 (green) localization and decreased intensity after MYC-AURA overexpression. Representative images (CAP350 in red, MYC in yellow) are shown in G (scale bar, 2 µm) and intensity quantification (normalized to CAP350) in H. (I and J) Examination of SCLT1 (green) localization and decreased intensity after MYC-AURA overexpression. Representative images (CAP350 in red, MYC in yellow) are shown in I (scale bar, 1 µm) and intensity quantification (normalized to CAP350) in J. (K and L) Examination of IFT57 (green) localization and decreased intensity at BBs after MYC-AURA overexpression. Representative images (Ac-tub in red, CAP350 in yellow) are shown in K (scale bar, 2 µm) and BB intensity quantification (normalized to CAP350) in L. Asterisks indicate statistical significance determined using an unpaired t test.

    Article Snippet: Next, following digest with AflII and BstBI (enzymes from NEB) the MYC-BirA* fragment was ligated (K1422; ThermoFisher) into AflII and BstBI sites of pgLAP2 vector (19703; Addgene).

    Techniques: Microscopy, Over Expression, Transfection

    Transcription of the master operon flhDC . ( A ) To verify the transcription of the plasmid-encoded master operon flhDC Reverse Transcriptase PCR Reaction of the isolated RNA was performed. It was assumed that within the overnight culture cells exhibiting flagellar assembly were found and therefore this culture was used as a positive control. Cells before addition of the inducer IPTG were used as reference compared to cells after induction of the plasmid-encoded flhDC . To evaluate genomic DNA contamination the isolated RNA samples were also subjected to subsequent PCR. Plasmid-encoded flhDC was used as a positive control for the PCR reaction. Fermentas 1 KB GeneRuler, ON overnight culture sample, -I whole cell sample without induction of plasmid-encoded flhDC , +I whole cell samples with induction of plasmid-encoded flhDC , RT Reverse Transcriptase Reaction, + plasmid-encoded flhDC ; ( B ) Scheme of the plasmid-encoded master operon under the control of the lacUV5 promoter

    Journal: PLoS ONE

    Article Title: FlgM as a Secretion Moiety for the Development of an Inducible Type III Secretion System

    doi: 10.1371/journal.pone.0059034

    Figure Lengend Snippet: Transcription of the master operon flhDC . ( A ) To verify the transcription of the plasmid-encoded master operon flhDC Reverse Transcriptase PCR Reaction of the isolated RNA was performed. It was assumed that within the overnight culture cells exhibiting flagellar assembly were found and therefore this culture was used as a positive control. Cells before addition of the inducer IPTG were used as reference compared to cells after induction of the plasmid-encoded flhDC . To evaluate genomic DNA contamination the isolated RNA samples were also subjected to subsequent PCR. Plasmid-encoded flhDC was used as a positive control for the PCR reaction. Fermentas 1 KB GeneRuler, ON overnight culture sample, -I whole cell sample without induction of plasmid-encoded flhDC , +I whole cell samples with induction of plasmid-encoded flhDC , RT Reverse Transcriptase Reaction, + plasmid-encoded flhDC ; ( B ) Scheme of the plasmid-encoded master operon under the control of the lacUV5 promoter

    Article Snippet: Rapid DNA ligation kit, Pfu DNA polymerase and 10× MgSO4 -PCR buffer were obtained from Fermentas.

    Techniques: Plasmid Preparation, Polymerase Chain Reaction, Isolation, Positive Control

    Expression of TxS in established prostate cancer cells. A: Western blot analysis of TxS expression in different PCa cell lines. TS denotes TxS. NHP, normal prostate epithelial cells. Platelet lysates as positive control. B: RT-PCR analysis of TxS mRNA expression. Shown is one round PCR product using first set of TxS specific primers as described in Materials and Methods. RT(-), RNA samples without reverse transcriptase were used as negative control to eliminate possible false positive results from genomic DNA contamination. C: Northern blot analysis of TxS expression. Left panel , ethidium bromide staining of RNA in agarose gels indicating equal loading of RNA samples. Right panel, autoradiograph of membrane probed with TxS cDNA probe. Lane 1 , RNA molecular weight markers; lane 2 , LNCaP; lane 3 , PC-3; lane 4 , DU145; lane 5 , HEL cells as positive control. The approximate size for TxS mRNA in PC-3 cells is 1.8 ∼1.9 kb.

    Journal: The American Journal of Pathology

    Article Title: Differential Expression of Thromboxane Synthase in Prostate Carcinoma

    doi:

    Figure Lengend Snippet: Expression of TxS in established prostate cancer cells. A: Western blot analysis of TxS expression in different PCa cell lines. TS denotes TxS. NHP, normal prostate epithelial cells. Platelet lysates as positive control. B: RT-PCR analysis of TxS mRNA expression. Shown is one round PCR product using first set of TxS specific primers as described in Materials and Methods. RT(-), RNA samples without reverse transcriptase were used as negative control to eliminate possible false positive results from genomic DNA contamination. C: Northern blot analysis of TxS expression. Left panel , ethidium bromide staining of RNA in agarose gels indicating equal loading of RNA samples. Right panel, autoradiograph of membrane probed with TxS cDNA probe. Lane 1 , RNA molecular weight markers; lane 2 , LNCaP; lane 3 , PC-3; lane 4 , DU145; lane 5 , HEL cells as positive control. The approximate size for TxS mRNA in PC-3 cells is 1.8 ∼1.9 kb.

    Article Snippet: The TOPO PCR cloning vector and DNA ligation kit were obtained from Invitrogen (Carlsbad, CA).

    Techniques: Expressing, Western Blot, Positive Control, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control, Northern Blot, Staining, Autoradiography, Molecular Weight