lr clonase enzyme  (Thermo Fisher)


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    Thermo Fisher lr clonase enzyme
    Lr Clonase Enzyme, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase enzyme/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    lr clonase enzyme - by Bioz Stars, 2020-02
    99/100 stars

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    Clone Assay:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Paragraph title: Cloning, stable, and transient plant transformation ... Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen).

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen). ..

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen). .. To generate LAF1 mutant genes (W87A or R97A), site-directed mutagenesis was performed using the QuikChange site-directed mutagenesis kit (Stratagene), pENTR-LAF1 as a template, and the following primer sets: 5′-TCTTCCTTGGGTAA CAAGGCGTCGCAAATAGCTAAATTC-3′ and 5′-GAATTT AGCTATTTGCGACGCCTTGTTACCCAAGGAAGA-3′ for LAF1(W87A), and 5′-GCTAAATTCTTACCGGGAGCAACAG ACAATGAGATAAAG-3′ and 5′-CTTTATCTCATTGTCTGT TGCTCCCGGTAAGAATTTAGC-3′ for LAF1(R97A).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: The complete sequence for the newly cloned SK-V isoform was deposited in GenBank (accession number MH001552). .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors. .. The coding regions of all cDNA constructs were sequenced using an ABI PRISM 3130 sequencer (Applied Biosystems Japan Ltd., Tokyo, Japan) and compared with the human gene ITGB1 (GenBank™ ) to confirm both the presence of the desired mutations ( ) and the absence of any additional mutations.

    Amplification:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Cloning, stable, and transient plant transformation The coding sequence of MP17 was fused C-terminally with six histidine residues by conventional PCR amplification (from plasmid p35S-1; Hofius et al., ; primer see Table ) and inserted in pBinAR (Höfgen and Willmitzer, ) via Asp718/BamHI to create plasmid MP17–PMH. .. Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen).

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: DNA sequences encoding these regions were amplified by PCR using full-length cDNAs of HFR1 ( ) and LAF1 ( ). cDNA encoding full-length PAT1 was amplified by PCR to generate pMBP-DC-PAT1. .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: For SK-M and SK-V cDNA amplification, PCR was performed using the reverse primer 5ʹ-TCAGCTAGAATGTGGAAACAGCAT-3ʹ, and either the forward primer 5ʹ-ATGTCAATTCA GAAGCTTAACGAC-3ʹ to target the SK-M isoform or the forward primer 5ʹ-ATGTCGCCGGCCTTCTGC-3ʹ to target the SK-V isoform. .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: CHO Lec 3.2.8.1 cells, a GnT-I-deficient cell line, were a gift from Dr. Pamela Stanly (Albert Einstein College of Medicine, New York) ( ). cDNA Constructs —The cDNA of integrin β1 subunit was amplified by PCR from the reverse-transcribed product of human placenta total RNA (OriGene Technologies, Inc., Rockville, MD) and then was inserted into a cloning vector (pENTR-D-Topo, Invitrogen), according to the manufacturer's protocol. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Plasmid Preparation:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: .. Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen). ..

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: The binary vector pBA002 ( ) for constitutive expression in plants was modified to obtain pHyg-DC, pBA-DC, pBA-6myc-DC, and pBA-DC-CFP. .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen).

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen). .. To generate LAF1 mutant genes (W87A or R97A), site-directed mutagenesis was performed using the QuikChange site-directed mutagenesis kit (Stratagene), pENTR-LAF1 as a template, and the following primer sets: 5′-TCTTCCTTGGGTAA CAAGGCGTCGCAAATAGCTAAATTC-3′ and 5′-GAATTT AGCTATTTGCGACGCCTTGTTACCCAAGGAAGA-3′ for LAF1(W87A), and 5′-GCTAAATTCTTACCGGGAGCAACAG ACAATGAGATAAAG-3′ and 5′-CTTTATCTCATTGTCTGT TGCTCCCGGTAAGAATTTAGC-3′ for LAF1(R97A).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies). .. CRISPR Gene Tagging A V5 epitope tag was introduced at the SK locus ( , and ) using a single-stranded oligodeoxynucloetide (ssODN) homology directed repair CRISPR-Cas9 approach ( ).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: The retrovirus vector, pBabe-Puro plasmid , was made to be Gateway-compatible using the Gateway Conversion System kit (Invitrogen), resulting in pBabe-puro-Rfa. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Article Title: The Involvement of the Banana F-Box Protein MaEBF1 in Regulating Chilling-Inhibited Starch Degradation through Interaction with a MaNAC67-Like Protein
    Article Snippet: Subcellular Location Analysis The full CDS of MaEBF1 and MaNAC67-like without stop codons were ligated into a pENTR/D vector (Invitrogen, Waltham, MA, USA). .. MaEBF1-GFP and MaNAC67-like-GFP fusion proteins were produced using the LR clonase enzyme (Invitrogen, USA).

    Polymerase Chain Reaction:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Different mutations were introduced in the MP17 sequence by overlap PCR with specific primers (Table ) or deletion variants were obtained by PCR with respective primers. .. Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen).

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: DNA sequences encoding these regions were amplified by PCR using full-length cDNAs of HFR1 ( ) and LAF1 ( ). cDNA encoding full-length PAT1 was amplified by PCR to generate pMBP-DC-PAT1. .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: The SK-M and SK-V PCR products were subsequently cloned into the TOPO-XL vector (Life Technologies) and fully sequenced. .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: CHO Lec 3.2.8.1 cells, a GnT-I-deficient cell line, were a gift from Dr. Pamela Stanly (Albert Einstein College of Medicine, New York) ( ). cDNA Constructs —The cDNA of integrin β1 subunit was amplified by PCR from the reverse-transcribed product of human placenta total RNA (OriGene Technologies, Inc., Rockville, MD) and then was inserted into a cloning vector (pENTR-D-Topo, Invitrogen), according to the manufacturer's protocol. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Mutagenesis:

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: Paragraph title: Vector construction and site-directed mutagenesis of LAF1 ... All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: Mutations were introduced into the β1 cDNA using a Quick-Change site-directed mutagenesis kit (Stratagene, La Jolla, CA.) according to the manufacturer's instructions. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Subcloning:

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies). .. CRISPR Gene Tagging A V5 epitope tag was introduced at the SK locus ( , and ) using a single-stranded oligodeoxynucloetide (ssODN) homology directed repair CRISPR-Cas9 approach ( ).

    Sequencing:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Different mutations were introduced in the MP17 sequence by overlap PCR with specific primers (Table ) or deletion variants were obtained by PCR with respective primers. .. Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: The complete sequence for the newly cloned SK-V isoform was deposited in GenBank (accession number MH001552). .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Knock-Out:

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: Cell Cultures —Epithelial GE11 cells, derived from β1 integrin knock-out embryonic stem cells , were maintained at 37 °C in Dulbecco's modified Eagle's medium (DMEM; Invitrogen), supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin, and 20 n m trichostatin A. Phoenix cells and integrin α5-deficient cells (CHO-B2) ( ) were grown in DMEM supplemented with 10% FBS, penicillin/streptomycin. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Produced:

    Article Title: The Involvement of the Banana F-Box Protein MaEBF1 in Regulating Chilling-Inhibited Starch Degradation through Interaction with a MaNAC67-Like Protein
    Article Snippet: .. MaEBF1-GFP and MaNAC67-like-GFP fusion proteins were produced using the LR clonase enzyme (Invitrogen, USA). .. The resulting plasmids and control GFP vector were then transformed in the Agrobacterium tumefaciens strain GV3101.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: For the cloning of SK-M and SK-V rescue constructs, RT-PCR was performed on total RNA extracts (TRIzol, Life Technologies) from Canton-S third instar larvae. .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Confocal Laser Scanning Microscopy:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen). .. Two days after infiltration, localization was analyzed by Confocal Laser Scanning Microscopy using a TCS SP5 (Leica; Bensheim).

    Construct:

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: The majority of constructs were made using the Gateway system (Invitrogen) ( ). .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen).

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: Most constructs except GST-tagged vectors in this study were made using the Gateway system (Invitrogen). .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen).

    Article Title: The Drosophila Small Conductance Calcium-Activated Potassium Channel Negatively Regulates Nociception
    Article Snippet: For the cloning of SK-M and SK-V rescue constructs, RT-PCR was performed on total RNA extracts (TRIzol, Life Technologies) from Canton-S third instar larvae. .. The following primers were used to target the desired SK-M and SK-V amplicons for subcloning into the pENTR/DTOPO vector (Thermo Fisher Scientific): 5ʹ-CACCATGTCAATTCAGAAGCTTAACGAC-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAG CATG-3ʹ for SK-M , or 5ʹ-CACCATGTCGCCGGCCTTCT-3ʹ and 5ʹ-TCAGCTAGAATGTGGAAACAGCATGGGC-3ʹ for SK-V. Gateway recombination technology was used to further subclone the SK-M and SK-V cDNAs from pENTR into the final pTW destination vector (i.e., pUASt-SK-M and pUASt-SK-V) via the LR Clonase enzyme (Life Technologies).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: CHO Lec 3.2.8.1 cells, a GnT-I-deficient cell line, were a gift from Dr. Pamela Stanly (Albert Einstein College of Medicine, New York) ( ). cDNA Constructs —The cDNA of integrin β1 subunit was amplified by PCR from the reverse-transcribed product of human placenta total RNA (OriGene Technologies, Inc., Rockville, MD) and then was inserted into a cloning vector (pENTR-D-Topo, Invitrogen), according to the manufacturer's protocol. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Expressing:

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: The vectors pER8 and pER10 ( ) for estradiol-inducible expression in plants under the control of the XVE system were modified to obtain pER8-6myc-DC and pER10-YFP-DC, respectively. .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen).

    Article Title: Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling
    Article Snippet: Plasmids encoding MBP-COP1, MBP-HFR1, and MBP-HFR1(C) for protein expression in Escherichia coli and plasmids encoding 35S-HFR1-3HA, 35S-HFR1-6Myc, and 35S-HFR1(C)-3HA for protein expression in plants were described previously ( ). .. All cDNA or DNA fragments were cloned in pENTR/D vector (Invitrogen) and then transferred into the appropriate vectors by recombination using the LR Clonase enzyme according to the manufacturer’s instructions (Invitrogen).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors. .. The soluble expression system of α5β1 integrin was clearly described by Takagi et al. ( , ).

    Modification:

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: The vectors pER8 and pER10 ( ) for estradiol-inducible expression in plants under the control of the XVE system were modified to obtain pER8-6myc-DC and pER10-YFP-DC, respectively. .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen).

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: Cell Cultures —Epithelial GE11 cells, derived from β1 integrin knock-out embryonic stem cells , were maintained at 37 °C in Dulbecco's modified Eagle's medium (DMEM; Invitrogen), supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin, and 20 n m trichostatin A. Phoenix cells and integrin α5-deficient cells (CHO-B2) ( ) were grown in DMEM supplemented with 10% FBS, penicillin/streptomycin. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

    Staining:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen). .. Propidium iodide staining and excitation/detection of emission were conducted as described (Vogel et al., ).

    Transformation Assay:

    Article Title: PD Trafficking of Potato Leaf Roll Virus Movement Protein in Arabidopsis Depends on Site-specific Protein Phosphorylation
    Article Snippet: Paragraph title: Cloning, stable, and transient plant transformation ... Mutagenized DNA fragments were first subcloned into the pENTR directional TOPO vector (Invitrogen) followed by LR reaction into the destination vector pK7FWG2 using LR clonase enzyme to create C-terminal GFP fusions (Invitrogen).

    Article Title: Cucumber mosaic virus-encoded 2b suppressor inhibits Arabidopsis Argonaute1 cleavage activity to counter plant defense
    Article Snippet: Several destination vectors (DC in our nomenclature) were created for transient expression in N. benthamiania and stable Arabidopsis transformation. .. Most of the cDNA or DNA fragments were cloned into pENTR/D vectors and then transferred to the appropriate vectors by recombination using the LR Clonase enzyme (Invitrogen).

    Article Title: The Involvement of the Banana F-Box Protein MaEBF1 in Regulating Chilling-Inhibited Starch Degradation through Interaction with a MaNAC67-Like Protein
    Article Snippet: MaEBF1-GFP and MaNAC67-like-GFP fusion proteins were produced using the LR clonase enzyme (Invitrogen, USA). .. The resulting plasmids and control GFP vector were then transformed in the Agrobacterium tumefaciens strain GV3101.

    Derivative Assay:

    Article Title: N-Glycosylation of the I-like Domain of ?1 Integrin Is Essential for ?1 Integrin Expression and Biological Function
    Article Snippet: Cell Cultures —Epithelial GE11 cells, derived from β1 integrin knock-out embryonic stem cells , were maintained at 37 °C in Dulbecco's modified Eagle's medium (DMEM; Invitrogen), supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin, and 20 n m trichostatin A. Phoenix cells and integrin α5-deficient cells (CHO-B2) ( ) were grown in DMEM supplemented with 10% FBS, penicillin/streptomycin. .. The LR clonase enzyme (Invitrogen) was used to transfer the cDNA of either β1 integrin or GFP from the cloning vectors.

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    Thermo Fisher lr clonase ii plus
    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite <t>Clonase</t> strategy utilizing modified MuLE entry vectors.
    Lr Clonase Ii Plus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher gateway lr clonase ii enzyme mix
    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite <t>Clonase</t> strategy utilizing modified MuLE entry vectors.
    Gateway Lr Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 286 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway lr clonase ii enzyme mix/product/Thermo Fisher
    Average 90 stars, based on 286 article reviews
    Price from $9.99 to $1999.99
    gateway lr clonase ii enzyme mix - by Bioz Stars, 2020-02
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    99
    Thermo Fisher lr clonase reaction
    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite <t>Clonase</t> strategy utilizing modified MuLE entry vectors.
    Lr Clonase Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase reaction/product/Thermo Fisher
    Average 99 stars, based on 169 article reviews
    Price from $9.99 to $1999.99
    lr clonase reaction - by Bioz Stars, 2020-02
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    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Journal: Gene

    Article Title: A Splice Junction-Targeted CRISPR Approach (spJCRISPR) Reveals Human FOXO3B To Be A Protein-Coding Gene

    doi: 10.1016/j.gene.2018.06.048

    Figure Lengend Snippet: FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Article Snippet: LR Clonase II Plus (Thermo 12538) reactions were performed with 2 μl Clonase LR II Plus, 20 fmoles pLenti X1 Puro DEST, 10 fmoles of each sgRNA pENTR and 10 fmoles of pENTR hCas9D10A all to 10 μl volume in Tris-EDTA (TE) buffer pH 8.

    Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Modification

    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Journal: Gene

    Article Title: A Splice Junction-Targeted CRISPR Approach (spJCRISPR) Reveals Human FOXO3B To Be A Protein-Coding Gene

    doi: 10.1016/j.gene.2018.06.048

    Figure Lengend Snippet: FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Article Snippet: LR Clonase II (Thermo 11791100) reactions were performed with a modified form of the pInducer20 vector (Addgene #44012) where the Neo resistance cassette was replaced with a blasticidin resistance cassette.

    Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Modification