in fusion hd cloning kit  (TaKaRa)

 
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    Name:
    In Fusion HD Cloning Kit
    Description:
    In Fusion Cloning offers a one step high efficiency and high throughput cloning method for projects that require a large number of cloning reactions such as expression screening studies and library construction
    Catalog Number:
    639650
    Price:
    None
    Size:
    100 Rxns
    Category:
    In Fusion Cloning for high throughput workflows In Fusion Cloning Cloning
    Buy from Supplier


    Structured Review

    TaKaRa in fusion hd cloning kit
    In Fusion Cloning offers a one step high efficiency and high throughput cloning method for projects that require a large number of cloning reactions such as expression screening studies and library construction
    https://www.bioz.com/result/in fusion hd cloning kit/product/TaKaRa
    Average 90 stars, based on 1032 article reviews
    Price from $9.99 to $1999.99
    in fusion hd cloning kit - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: .. Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter. .. Rat IL-10 was PCR cloned from rat splenocyte cDNA, as previously described.

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio). ..

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB). ..

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: .. The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S. .. The two fragments were ligated using the In-Fusion HD Cloning Kit.

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally epitope-tagged full-length Drosophila Kat80 were created by PCR amplification of the CDS of full-length Kat80 (amino acids 1–819) from the BDGP cDNA clone LD44201 (Drosophila Genomics Resource Center) with the CDS of Myc or FLAG engineered onto the 5’ end of the PCR product.

    Article Title: Crystal structures of the Arabidopsis thaliana organellar RNA editing factors MORF1 and MORF9
    Article Snippet: .. The pMAL-TEV-MCS41 vector was created as follows: pET-41-a (Novagen) multiple cloning site between SacI to XhoI was amplified by PCR and cloned into SacI –HindIII sites into the pMAL-TEV vector by the In-Fusion® HD Cloning kit (TAKARA). .. The pET41-GFP vector was created as follows: GFP CDS from the pSM-GFP (GenBank: U70495.1) was amplified and cloned into the pET-41-a (Novagen) NotI site by the In-Fusion® HD Cloning kit.

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). ..

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Article Title: Cloning, localization and focus formation at DNA damage sites of canine Ku70
    Article Snippet: .. Each cycle consisted of a denaturation step at 94°C for 0.5 min, annealing at 60°C for 0.5 min and extension at 72°C for 2 min. PCR products were subcloned into the Eco RI and Apa I sites of pEYFP-C1 (pEYFP-canine Ku70 ) using the In-Fusion HD cloning kit (Takara Bio Inc.), and the inserts were validated by sequencing. .. Other PCR primers used in this study were as follows: dX6 Nseq F1: 5′-GCACAAGGGTGAGCAATAAACAATATG-3′, dX6 Nseq R1: 5′-TGGATACACTGGATGCTCATGTCAAAAG-3′, dX6 Cseq F2: 5′-GGTGGGTTGAAGAAGCAGGAG-3′ and dX6 Cseq R2: 5-AAAGCACAGCAAAGTGGGAGG-3′.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions. .. For constructing the PLK1 shRNA-expression plasmid vector, the DNA fragment containing a PLK1 shRNA coding region was cloned into BamHI/SpeI sites of pTRE3G1.

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: .. The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2. .. The URA3 marker in pLUMCL2 was then replaced with the NAT1 marker to generate the NAT1-Clox cassette in the plasmid pLNMCL.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: .. Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The DNA fragment corresponding to AM1_1557g2 was PCR amplified using the synthetic primers 5′-CGCGGCAGCCATATGTATGAGCGTAATATTGCT-3′ (forward primer) and 5′-CTCGAATTCGGATCCTCATGCTTCTGCTTTATCTCT-3′ (reverse primer), genomic DNA from A. marina MBIC11017, and PrimeSTAR Max DNA polymerase. pET28a was PCR amplified using the synthetic primers 5′-CATATGGCTGCCGCGCGG-3′ (forward primer) and 5′-GGATCCGAATTCGAGCTC-3′ (reverse primer), pET28a, and PrimeSTAR Max DNA polymerase.

    Article Title: Construction of a synthetic metabolic pathway for biosynthesis of the non-natural methionine precursor 2,4-dihydroxybutyric acid
    Article Snippet: .. Construction of pDHB operon: The three genes, Ec-lysC V115A:E119S:E250K:E343V , Bs-asd E218Q and Ms-ssr H39R N43H were cloned into the medium-copy pACT3 vector using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.) and following the user manual instructions. ..

    Amplification:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: For the targeting vector, 5′ and 3′ homology arms (0.71 kb and 0.82 kb, respectively) of PAX2 were amplified from the genomic DNA of iPS cells with the following primers: 5′-TGAATTCTTAGAGAGACACACACCGGG-3′ and 5′-TGCTAGCGGGGAGGCAGAGGAGCGGGA-3′ for the 5′ homology arm, and 5′-AACCTAGATCGGATCTGCACCGTGAGTACCGGCGC-3′ and 5′-ATTACGCCAAGCTTGCTGGCTCTCTCCCTGACTTC-3′ for the 3′ homology arm. .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB). ..

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: A GFP sequence that includes the promoter Ptet was amplified from the pBRgalKGR plasmid (a pBR322 derivative) [ , ] using the pBR_gfp_kanR_Fw and pBR_gfp_kanR_Rv primers. .. The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S.

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally epitope-tagged full-length Drosophila Kat80 were created by PCR amplification of the CDS of full-length Kat80 (amino acids 1–819) from the BDGP cDNA clone LD44201 (Drosophila Genomics Resource Center) with the CDS of Myc or FLAG engineered onto the 5’ end of the PCR product.

    Article Title: Crystal structures of the Arabidopsis thaliana organellar RNA editing factors MORF1 and MORF9
    Article Snippet: .. The pMAL-TEV-MCS41 vector was created as follows: pET-41-a (Novagen) multiple cloning site between SacI to XhoI was amplified by PCR and cloned into SacI –HindIII sites into the pMAL-TEV vector by the In-Fusion® HD Cloning kit (TAKARA). .. The pET41-GFP vector was created as follows: GFP CDS from the pSM-GFP (GenBank: U70495.1) was amplified and cloned into the pET-41-a (Novagen) NotI site by the In-Fusion® HD Cloning kit.

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: A 1681 bp fragment of the human ABCC4 gene was amplified from the genomic DNA of −1508GG and −1508AA homozygous subjects by PCR using the following primers: (forward) 5′- TCTATCGATAGGTAC GGCCATGCTTAGACATAGGCTTA-3′ and (reverse) 5′- GATCGCAGATCTCGA AGAACACGCGTGAGCAGAGGTT-3′. .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA).

    Article Title: Cloning, localization and focus formation at DNA damage sites of canine Ku70
    Article Snippet: This was followed by 40 cycles of PCR amplification. .. Each cycle consisted of a denaturation step at 94°C for 0.5 min, annealing at 60°C for 0.5 min and extension at 72°C for 2 min. PCR products were subcloned into the Eco RI and Apa I sites of pEYFP-C1 (pEYFP-canine Ku70 ) using the In-Fusion HD cloning kit (Takara Bio Inc.), and the inserts were validated by sequencing.

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2. .. NAT1 was amplified from pJK863 using the primers Clox-NAT1-F and Clox-NAT1-R , and then cloned between the Bpu10I and NheI sites of pLUMCL2 by In-Fusion cloning to create pLNMCL.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The DNA fragment corresponding to AM1_1557g2 was PCR amplified using the synthetic primers 5′-CGCGGCAGCCATATGTATGAGCGTAATATTGCT-3′ (forward primer) and 5′-CTCGAATTCGGATCCTCATGCTTCTGCTTTATCTCT-3′ (reverse primer), genomic DNA from A. marina MBIC11017, and PrimeSTAR Max DNA polymerase. pET28a was PCR amplified using the synthetic primers 5′-CATATGGCTGCCGCGCGG-3′ (forward primer) and 5′-GGATCCGAATTCGAGCTC-3′ (reverse primer), pET28a, and PrimeSTAR Max DNA polymerase.

    Mass Spectrometry:

    Article Title: Construction of a synthetic metabolic pathway for biosynthesis of the non-natural methionine precursor 2,4-dihydroxybutyric acid
    Article Snippet: .. Construction of pDHB operon: The three genes, Ec-lysC V115A:E119S:E250K:E343V , Bs-asd E218Q and Ms-ssr H39R N43H were cloned into the medium-copy pACT3 vector using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.) and following the user manual instructions. ..

    Synthesized:

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions. .. For constructing the PLK1 shRNA-expression plasmid vector, the DNA fragment containing a PLK1 shRNA coding region was cloned into BamHI/SpeI sites of pTRE3G1.

    Construct:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: Plasmid construction Multi-expression plasmids were constructed in the pMT/V5-His A expression plasmid backbone (Invitrogen) and additional expression cassettes were cloned into unique restriction enzyme sites engineered into regions outside of the copper-inducible expression cassette. .. The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: Plasmids The constitutive, human codon-optimized, Streptococcus pyogenes Cas9 retroviral expression construct (MSCV-hCas9-PGK-Puro, Addgene: #65655) and lentiviral sgRNA expression vector (LRG, Addgene: #65656) were adapted from previous work [ ]. .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909).

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: The ApaI/XbaI region of rrnB operon was subcloned between HindIII and EcoRI sites of pUC18 in order to facilitate the generation of Spinach ribosome constructs (pUC18-AX). .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB).

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. The multi-expression plasmids constructed in this study are shown in .

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: Paragraph title: 2.5. Activity of the ABCC4 Promoter Constructs ... PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA).

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The pTRE3G1 plasmid vector contains the following constructs: reverse tetracycline activator coding region derived from pCMV-TET3G (Takara Bio Inc., Shiga, Japan); an improved variant of the copepod Pontellina plumata green fluorescent protein (max GFP) coding region derived from pmaxGFP (Lonza, Basel, Switzerland); the GAPDH promoter from nucleotides –376 to + 183 relative to the transcription start site; a tetracycline-response element promoter derived from pTRE3G (Takara Bio); 5′ and 3′ miR-155 flunking region derived from pcDNA 6.2-GW/EmGFP-miR (Thermo Fisher Scientific); and a Luciferase shRNA coding region derived from pSingle-tTS-Anti-Luc (Takara Bio Inc.) for control shRNA. .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: Clox Construction A synthetic, codon-optimized cre open reading frame, interrupted by a C. albicans TUB2 intron at codon 135, was designed in silico (Results), constructed by DNA2.0 (Menlo Park, CA, USA) and cloned between the Nhe I and Nco I sites in pLUL2 to generate pLUCL2. .. The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. A plasmid expressing AM1_1557g2 (pET28a_AM1_1557g2) was then constructed with the TaKaRa in-fusion system reagents. pET28a_AM1_1557g2_C304A was generated using the primers 5′-AGAGACGCACATTTAGAGATTTTGGAA-3′ (forward primer) and 5′-TAAATGTGCGTCTCTATAAGATTCTTG-3′ (reverse primer), pET28a_AM1_1557g2, and PrimeSTAR Max Basal Mutagenesis kit reagents (TaKaRa). pET28a_AM1_1557g2_L337N was obtained in the same way as described above with a primer set (5′- CTATCAGAATAACGTCCCACGTCAATG-3′, 5′- ACGTTATTCTGATAGGCTGCCAGCAA-3′).

    Article Title: Construction of a synthetic metabolic pathway for biosynthesis of the non-natural methionine precursor 2,4-dihydroxybutyric acid
    Article Snippet: Construction plasmids for biosynthesis of DHB All plasmids constructed and used in this study are listed in . .. Construction of pDHB operon: The three genes, Ec-lysC V115A:E119S:E250K:E343V , Bs-asd E218Q and Ms-ssr H39R N43H were cloned into the medium-copy pACT3 vector using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.) and following the user manual instructions.

    Real-time Polymerase Chain Reaction:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter. .. AAV vector titer was determined by real-time PCR using a 7500 Fast Real-Time Instrument (Applied Biosystems) with rat IL-10 gene primers: forward, 5′-TAAGCTCCAAGACAAAGGGTG-3′; and reverse, 5′-GTCCTCCAGTCCAGTAGATG-3′.

    Luciferase:

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The pTRE3G1 plasmid vector contains the following constructs: reverse tetracycline activator coding region derived from pCMV-TET3G (Takara Bio Inc., Shiga, Japan); an improved variant of the copepod Pontellina plumata green fluorescent protein (max GFP) coding region derived from pmaxGFP (Lonza, Basel, Switzerland); the GAPDH promoter from nucleotides –376 to + 183 relative to the transcription start site; a tetracycline-response element promoter derived from pTRE3G (Takara Bio); 5′ and 3′ miR-155 flunking region derived from pcDNA 6.2-GW/EmGFP-miR (Thermo Fisher Scientific); and a Luciferase shRNA coding region derived from pSingle-tTS-Anti-Luc (Takara Bio Inc.) for control shRNA. .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Activity Assay:

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: The PAX2 -TALEN plasmids were transfected into HEK293 cells using Lipofectamine 2000 (Thermo Fisher Scientific) to evaluate their activity. .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: Paragraph title: 2.5. Activity of the ABCC4 Promoter Constructs ... PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA).

    In Silico:

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: Clox Construction A synthetic, codon-optimized cre open reading frame, interrupted by a C. albicans TUB2 intron at codon 135, was designed in silico (Results), constructed by DNA2.0 (Menlo Park, CA, USA) and cloned between the Nhe I and Nco I sites in pLUL2 to generate pLUCL2. .. The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2.

    Expressing:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: All copper-inducible transgenes were created by cloning PCR-amplified CDS into the multiple cloning site of the copper-inducible expression cassette. .. The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The wild-type human DOT1L cDNA was cloned into a lentiviral expression vector with EFS prompter and P2A-linked Puromycin resistance gene. .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909).

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. A plasmid expressing AM1_1557g2 (pET28a_AM1_1557g2) was then constructed with the TaKaRa in-fusion system reagents. pET28a_AM1_1557g2_C304A was generated using the primers 5′-AGAGACGCACATTTAGAGATTTTGGAA-3′ (forward primer) and 5′-TAAATGTGCGTCTCTATAAGATTCTTG-3′ (reverse primer), pET28a_AM1_1557g2, and PrimeSTAR Max Basal Mutagenesis kit reagents (TaKaRa). pET28a_AM1_1557g2_L337N was obtained in the same way as described above with a primer set (5′- CTATCAGAATAACGTCCCACGTCAATG-3′, 5′- ACGTTATTCTGATAGGCTGCCAGCAA-3′).

    Modification:

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: Upon sequence verification, the 5′ homology arm was cloned in the EcoRI-NheI site of the modified HR120PA-1 vector (Systems Biosciences), such that the start codon of PAX2 was replaced with that of TurboGFP , followed by red fluorescent protein (RFP ) and puromycin-resistance gene (Puro ) cassettes flanked by loxP and insulator sequences. .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: Activity of the ABCC4 Promoter Constructs Human mast cells (HMC-1) were cultured in Iscove's Modified Dulbecco's Medium (Gibco, Grand Island, NY, USA). .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA).

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Derivative Assay:

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: Thus, all three plasmids that were produced had a common site of initiation for replication derived from pUC19, and carried a single gene for ampicillin resistance. .. The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The pTRE3G1 plasmid vector contains the following constructs: reverse tetracycline activator coding region derived from pCMV-TET3G (Takara Bio Inc., Shiga, Japan); an improved variant of the copepod Pontellina plumata green fluorescent protein (max GFP) coding region derived from pmaxGFP (Lonza, Basel, Switzerland); the GAPDH promoter from nucleotides –376 to + 183 relative to the transcription start site; a tetracycline-response element promoter derived from pTRE3G (Takara Bio); 5′ and 3′ miR-155 flunking region derived from pcDNA 6.2-GW/EmGFP-miR (Thermo Fisher Scientific); and a Luciferase shRNA coding region derived from pSingle-tTS-Anti-Luc (Takara Bio Inc.) for control shRNA. .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Transfection:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter. .. Recombinant dsAAV vectors (dsAAV1-CAG-IL-10) were generated from cultured serum-free medium using polyethylenimine-based triple transfections of HEK293 cells.

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: The PAX2 -TALEN plasmids were transfected into HEK293 cells using Lipofectamine 2000 (Thermo Fisher Scientific) to evaluate their activity. .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). .. The constructs were transfected into A549, HMC-1, and U937 cells using Lipofectamine (Invitrogen) according to the manufacturer's protocol.

    Ligation:

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB). ..

    Cell Culture:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter. .. Recombinant dsAAV vectors (dsAAV1-CAG-IL-10) were generated from cultured serum-free medium using polyethylenimine-based triple transfections of HEK293 cells.

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: A549, human alveolar type II epithelial-like, and U937, human leukemic monocyte lymphoma cell lines, were cultured in Roswell Park Memorial Institute-1640 Medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin G sodium, and 100 μ g/mL streptomycin sulfate (Gibco) at 37°C in a 5% CO2 incubator. .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA).

    Generated:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Plasmid and Vector Construction The recombinant double-strand AAV vector plasmid pdsAAV-CBA-EGFP was a kind gift from Arun Srivastava (University of Florida College of Medicine). pdsAAV-CAG-EGFP was generated by replacing the chicken beta-actin (CBA) promoter region of pdsAAV-CBA-EGFP with the cytomegalovirus immediate-early enhancer/chicken β-actin hybrid (CAG) promoter region, as previously described. .. Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter.

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: Plasmid construction and production of PLK1 -specific shRNA and siRNA The doxycycline-regulated and RNA polymerase II-inducible shRNA-expression plasmid vector, designated as pTRE3G1, was generated for this study. .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. A plasmid expressing AM1_1557g2 (pET28a_AM1_1557g2) was then constructed with the TaKaRa in-fusion system reagents. pET28a_AM1_1557g2_C304A was generated using the primers 5′-AGAGACGCACATTTAGAGATTTTGGAA-3′ (forward primer) and 5′-TAAATGTGCGTCTCTATAAGATTCTTG-3′ (reverse primer), pET28a_AM1_1557g2, and PrimeSTAR Max Basal Mutagenesis kit reagents (TaKaRa). pET28a_AM1_1557g2_L337N was obtained in the same way as described above with a primer set (5′- CTATCAGAATAACGTCCCACGTCAATG-3′, 5′- ACGTTATTCTGATAGGCTGCCAGCAA-3′).

    DNA Sequencing:

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The sequences of the genes encoding AM1_1557g2, C304A and L337N were verified by DNA sequencing.

    Sequencing:

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: Upon sequence verification, the 5′ homology arm was cloned in the EcoRI-NheI site of the modified HR120PA-1 vector (Systems Biosciences), such that the start codon of PAX2 was replaced with that of TurboGFP , followed by red fluorescent protein (RFP ) and puromycin-resistance gene (Puro ) cassettes flanked by loxP and insulator sequences. .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB). ..

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: A GFP sequence that includes the promoter Ptet was amplified from the pBRgalKGR plasmid (a pBR322 derivative) [ , ] using the pBR_gfp_kanR_Fw and pBR_gfp_kanR_Rv primers. .. The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S.

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. To generate GEN1nuc , a 3xNLS sequence (DPKKKRKVDPKKKRKVDPKKKRKV) and a 3xFLAG tag sequence (AGDYKDHDGDYKDHIDYKDDDDK) were added to the C terminus of GEN1 by PCR.

    Article Title: Cloning, localization and focus formation at DNA damage sites of canine Ku70
    Article Snippet: .. Each cycle consisted of a denaturation step at 94°C for 0.5 min, annealing at 60°C for 0.5 min and extension at 72°C for 2 min. PCR products were subcloned into the Eco RI and Apa I sites of pEYFP-C1 (pEYFP-canine Ku70 ) using the In-Fusion HD cloning kit (Takara Bio Inc.), and the inserts were validated by sequencing. .. Other PCR primers used in this study were as follows: dX6 Nseq F1: 5′-GCACAAGGGTGAGCAATAAACAATATG-3′, dX6 Nseq R1: 5′-TGGATACACTGGATGCTCATGTCAAAAG-3′, dX6 Cseq F2: 5′-GGTGGGTTGAAGAAGCAGGAG-3′ and dX6 Cseq R2: 5-AAAGCACAGCAAAGTGGGAGG-3′.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions. .. The PLK1 shRNA sequence was designed using siDirect version 2.0 software ( http://sidirect2.rnai.jp/ ).

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: .. Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The DNA fragment corresponding to AM1_1557g2 was PCR amplified using the synthetic primers 5′-CGCGGCAGCCATATGTATGAGCGTAATATTGCT-3′ (forward primer) and 5′-CTCGAATTCGGATCCTCATGCTTCTGCTTTATCTCT-3′ (reverse primer), genomic DNA from A. marina MBIC11017, and PrimeSTAR Max DNA polymerase. pET28a was PCR amplified using the synthetic primers 5′-CATATGGCTGCCGCGCGG-3′ (forward primer) and 5′-GGATCCGAATTCGAGCTC-3′ (reverse primer), pET28a, and PrimeSTAR Max DNA polymerase.

    Article Title: Construction of a synthetic metabolic pathway for biosynthesis of the non-natural methionine precursor 2,4-dihydroxybutyric acid
    Article Snippet: Construction of pDHB operon: The three genes, Ec-lysC V115A:E119S:E250K:E343V , Bs-asd E218Q and Ms-ssr H39R N43H were cloned into the medium-copy pACT3 vector using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.) and following the user manual instructions. .. The forward primer inserted a unique restriction site and RBS in front of the gene of interest, and the reverse primer had a 15 bp overlapping sequence with the next forward primer. lysC-IF-forward and ssr-IF-reverse primers had 15 bp of sequence complementarity with the 5′ and 3′ ends of the plasmid, respectively.

    Affinity Purification:

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Recombinant:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Plasmid and Vector Construction The recombinant double-strand AAV vector plasmid pdsAAV-CBA-EGFP was a kind gift from Arun Srivastava (University of Florida College of Medicine). pdsAAV-CAG-EGFP was generated by replacing the chicken beta-actin (CBA) promoter region of pdsAAV-CBA-EGFP with the cytomegalovirus immediate-early enhancer/chicken β-actin hybrid (CAG) promoter region, as previously described. .. Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter.

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Crocin Bleaching Assay:

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Plasmid and Vector Construction The recombinant double-strand AAV vector plasmid pdsAAV-CBA-EGFP was a kind gift from Arun Srivastava (University of Florida College of Medicine). pdsAAV-CAG-EGFP was generated by replacing the chicken beta-actin (CBA) promoter region of pdsAAV-CBA-EGFP with the cytomegalovirus immediate-early enhancer/chicken β-actin hybrid (CAG) promoter region, as previously described. .. Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter.

    Mutagenesis:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. All point mutations were introduced by a PCR-based site-directed mutagenesis method and all plasmids were fully sequenced.

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: Both the DOT1L VVEL293MM DOT1L and EZH2 TR683KK mutations were introduced by standard PCR mutagenesis. .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909).

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. A plasmid expressing AM1_1557g2 (pET28a_AM1_1557g2) was then constructed with the TaKaRa in-fusion system reagents. pET28a_AM1_1557g2_C304A was generated using the primers 5′-AGAGACGCACATTTAGAGATTTTGGAA-3′ (forward primer) and 5′-TAAATGTGCGTCTCTATAAGATTCTTG-3′ (reverse primer), pET28a_AM1_1557g2, and PrimeSTAR Max Basal Mutagenesis kit reagents (TaKaRa). pET28a_AM1_1557g2_L337N was obtained in the same way as described above with a primer set (5′- CTATCAGAATAACGTCCCACGTCAATG-3′, 5′- ACGTTATTCTGATAGGCTGCCAGCAA-3′).

    Electrophoretic Mobility Shift Assay:

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: When the resulting fragment was denatured and annealed, a clear band shift was observed, indicating the formation of a mismatched duplex resulting from deletions or insertions . .. The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Purification:

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). ..

    Polymerase Chain Reaction:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter. .. Rat IL-10 was PCR cloned from rat splenocyte cDNA, as previously described.

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: .. Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB). ..

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: .. Transgenes encoding NH2 -terminally GFP-tagged deletion mutants of Kat60 were created by PCR amplification of the CDS of GFP and Kat60-ΔMIT or Kat60-AAA and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.). .. Transgenes encoding NH2 -terminally epitope-tagged full-length Drosophila Kat80 were created by PCR amplification of the CDS of full-length Kat80 (amino acids 1–819) from the BDGP cDNA clone LD44201 (Drosophila Genomics Resource Center) with the CDS of Myc or FLAG engineered onto the 5’ end of the PCR product.

    Article Title: Crystal structures of the Arabidopsis thaliana organellar RNA editing factors MORF1 and MORF9
    Article Snippet: .. The pMAL-TEV-MCS41 vector was created as follows: pET-41-a (Novagen) multiple cloning site between SacI to XhoI was amplified by PCR and cloned into SacI –HindIII sites into the pMAL-TEV vector by the In-Fusion® HD Cloning kit (TAKARA). .. The pET41-GFP vector was created as follows: GFP CDS from the pSM-GFP (GenBank: U70495.1) was amplified and cloned into the pET-41-a (Novagen) NotI site by the In-Fusion® HD Cloning kit.

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). ..

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Article Title: Cloning, localization and focus formation at DNA damage sites of canine Ku70
    Article Snippet: .. Each cycle consisted of a denaturation step at 94°C for 0.5 min, annealing at 60°C for 0.5 min and extension at 72°C for 2 min. PCR products were subcloned into the Eco RI and Apa I sites of pEYFP-C1 (pEYFP-canine Ku70 ) using the In-Fusion HD cloning kit (Takara Bio Inc.), and the inserts were validated by sequencing. .. Other PCR primers used in this study were as follows: dX6 Nseq F1: 5′-GCACAAGGGTGAGCAATAAACAATATG-3′, dX6 Nseq R1: 5′-TGGATACACTGGATGCTCATGTCAAAAG-3′, dX6 Cseq F2: 5′-GGTGGGTTGAAGAAGCAGGAG-3′ and dX6 Cseq R2: 5-AAAGCACAGCAAAGTGGGAGG-3′.

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: .. The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2. .. The URA3 marker in pLUMCL2 was then replaced with the NAT1 marker to generate the NAT1-Clox cassette in the plasmid pLNMCL.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The DNA fragment corresponding to AM1_1557g2 was PCR amplified using the synthetic primers 5′-CGCGGCAGCCATATGTATGAGCGTAATATTGCT-3′ (forward primer) and 5′-CTCGAATTCGGATCCTCATGCTTCTGCTTTATCTCT-3′ (reverse primer), genomic DNA from A. marina MBIC11017, and PrimeSTAR Max DNA polymerase. pET28a was PCR amplified using the synthetic primers 5′-CATATGGCTGCCGCGCGG-3′ (forward primer) and 5′-GGATCCGAATTCGAGCTC-3′ (reverse primer), pET28a, and PrimeSTAR Max DNA polymerase.

    Positron Emission Tomography:

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909). .. For the recombinant DOT1L KMT domain, wild-type and VVEL293MM mutant cDNA were cloned into the bacterial expression vector pET-22b (Novagen) via sequence- and ligation-Independent cloning, which contained an N-terminal His6 -tag and TEV protease site that allowed for affinity purification and subsequent cleavage of the affinity tag.

    Article Title: Crystal structures of the Arabidopsis thaliana organellar RNA editing factors MORF1 and MORF9
    Article Snippet: .. The pMAL-TEV-MCS41 vector was created as follows: pET-41-a (Novagen) multiple cloning site between SacI to XhoI was amplified by PCR and cloned into SacI –HindIII sites into the pMAL-TEV vector by the In-Fusion® HD Cloning kit (TAKARA). .. The pET41-GFP vector was created as follows: GFP CDS from the pSM-GFP (GenBank: U70495.1) was amplified and cloned into the pET-41-a (Novagen) NotI site by the In-Fusion® HD Cloning kit.

    cDNA Library Assay:

    Article Title: Cloning, localization and focus formation at DNA damage sites of canine Ku70
    Article Snippet: Cloning of canine Ku70 Oligonucleotide primers used to amplify canine Ku70 cDNA from a male beagle dog cDNA library (Biochain, Newark, CA, U.S.A.) were designed based on the predicted Ku70 genomic sequence of a female boxer dog, Canis lupus familiaris (XM_531714.3). .. Each cycle consisted of a denaturation step at 94°C for 0.5 min, annealing at 60°C for 0.5 min and extension at 72°C for 2 min. PCR products were subcloned into the Eco RI and Apa I sites of pEYFP-C1 (pEYFP-canine Ku70 ) using the In-Fusion HD cloning kit (Takara Bio Inc.), and the inserts were validated by sequencing.

    Plasmid Preparation:

    Article Title: The Non-Catalytic Domains of Drosophila Katanin Regulate Its Abundance and Microtubule-Disassembly Activity
    Article Snippet: Paragraph title: Plasmid construction ... The transgene encoding internally GFP-tagged full-length Kat60 was created by PCR amplification of the CDS of Kat60-MIT (amino acids 1–96), GFP, and Kat60-ΔMIT and fusion of the resulting PCR products using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.).

    Article Title: Mesenchymal Stem Cells Overexpressing Interleukin-10 Promote Neuroprotection in Experimental Acute Ischemic Stroke
    Article Snippet: Paragraph title: Plasmid and Vector Construction ... Using the In-Fusion HD Cloning Kit (TakaRa Bio), the EGFP region of pdsAAV-CAG-EGFP was replaced with rat IL-10 to generate pdsAAV-CAG-IL-10: rat IL-10 driven by the CAG promoter.

    Article Title: Rapid generation of drug-resistance alleles at endogenous loci using CRISPR-Cas9 indel mutagenesis
    Article Snippet: The wild-type EHZ2 cDNA was cloned into an MSCV-based vector containing a puromycin-resistance gene and a GFP reporter as previously reported [ ]. .. The PCR cloning procedures were performed using the In-Fusion® HD Cloning Kit (Clontech: #638909).

    Article Title: PAX2 is dispensable for in vitro nephron formation from human induced pluripotent stem cells
    Article Snippet: Paragraph title: Generation of PAX2-TALEN plasmids and the targeting vector ... The 3′ homology arm was cloned in the BamHI-SphI site using an In-fusion HD Cloning Kit (Takara Bio).

    Article Title: Use of Baby Spinach and Broccoli for imaging of structured cellular RNAs
    Article Snippet: Paragraph title: Plasmid construction ... Each Spinach sequence was amplified by PCR and cloned into pUC18-AXP using either an In-Fusion HD Cloning Kit (Clontech) or a Quick Ligation Kit (NEB).

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: Paragraph title: Plasmid construction ... The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S.

    Article Title: Crystal structures of the Arabidopsis thaliana organellar RNA editing factors MORF1 and MORF9
    Article Snippet: .. The pMAL-TEV-MCS41 vector was created as follows: pET-41-a (Novagen) multiple cloning site between SacI to XhoI was amplified by PCR and cloned into SacI –HindIII sites into the pMAL-TEV vector by the In-Fusion® HD Cloning kit (TAKARA). .. The pET41-GFP vector was created as follows: GFP CDS from the pSM-GFP (GenBank: U70495.1) was amplified and cloned into the pET-41-a (Novagen) NotI site by the In-Fusion® HD Cloning kit.

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). ..

    Article Title: Spatial control of the GEN1 Holliday junction resolvase ensures genome stability
    Article Snippet: .. Plasmids The GEN1 truncation constructs (full-length GEN1, GEN11–220 , GEN1221–444 , GEN1444–527 , GEN1526–662 , GEN1526–683 and GEN1662–908 ) were generated by PCR and cloned in-frame into a modified pDONR221 vector (Life Technologies) encoding a C-terminal GFP tag using an In-Fusion HD cloning kit (Clontech). .. The coding sequences were then shuttled by Gateway recombination (Life Technologies) into a modified pcDNA5/FRT vector (Life Technologies) with a Gateway cassette RfB cloned into the EcoRV site.

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: Paragraph title: Plasmid construction and production of PLK1 -specific shRNA and siRNA ... The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: .. The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2. .. The URA3 marker in pLUMCL2 was then replaced with the NAT1 marker to generate the NAT1-Clox cassette in the plasmid pLNMCL.

    Article Title: A biliverdin-binding cyanobacteriochrome from the chlorophyll d–bearing cyanobacterium Acaryochloris marina
    Article Snippet: .. Plasmid Construction The nucleotide sequence of AM1_1557g2 was cloned into pET28a (Novagen) using the In-Fusion HD Cloning kit (TaKaRa). .. The DNA fragment corresponding to AM1_1557g2 was PCR amplified using the synthetic primers 5′-CGCGGCAGCCATATGTATGAGCGTAATATTGCT-3′ (forward primer) and 5′-CTCGAATTCGGATCCTCATGCTTCTGCTTTATCTCT-3′ (reverse primer), genomic DNA from A. marina MBIC11017, and PrimeSTAR Max DNA polymerase. pET28a was PCR amplified using the synthetic primers 5′-CATATGGCTGCCGCGCGG-3′ (forward primer) and 5′-GGATCCGAATTCGAGCTC-3′ (reverse primer), pET28a, and PrimeSTAR Max DNA polymerase.

    Article Title: Construction of a synthetic metabolic pathway for biosynthesis of the non-natural methionine precursor 2,4-dihydroxybutyric acid
    Article Snippet: .. Construction of pDHB operon: The three genes, Ec-lysC V115A:E119S:E250K:E343V , Bs-asd E218Q and Ms-ssr H39R N43H were cloned into the medium-copy pACT3 vector using the In-Fusion HD cloning kit (Clontech Laboratories, Inc.) and following the user manual instructions. ..

    Software:

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions. .. The PLK1 shRNA sequence was designed using siDirect version 2.0 software ( http://sidirect2.rnai.jp/ ).

    shRNA:

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: Paragraph title: Plasmid construction and production of PLK1 -specific shRNA and siRNA ... The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

    Agarose Gel Electrophoresis:

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: .. PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). ..

    Spectrophotometry:

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). .. Plasmid DNAs were prepared from these constructs using the Endo-Free Plasmid Maxi Kit (Qiagen, Hilden, Germany), and the concentration and purity were assessed by UV spectrophotometry and agarose gel electrophoresis.

    Produced:

    Article Title: A reduced genome decreases the host carrying capacity for foreign DNA
    Article Snippet: Thus, all three plasmids that were produced had a common site of initiation for replication derived from pUC19, and carried a single gene for ampicillin resistance. .. The lacZ region of pUC19 was also removed using the In-Fusion HD Cloning Kit (Clontech) but with a different primer set (pUC19_del_lacZ_linearize_Fw and pUC19_del_lacZ_linearize_Rv2) , resulting in a 2,334-bp-long fragment, the same length as pUC-S.

    Concentration Assay:

    Article Title: A Role of the ABCC4 Gene Polymorphism in Airway Inflammation of Asthmatics
    Article Snippet: PCR products were gel purified using an Agarose Gel Purification Kit (GeneAll Biotechnology, Seoul, Korea) and ligated into the pGL3-basic vector (Promega, Madison, WI, USA) after digestion with KpnІ and XhoІ (Takara, Shuzo, Japan) using the In-Fusion® HD Cloning Kit (Clontech Laboratories Inc., Mountain View, CA, USA). .. Plasmid DNAs were prepared from these constructs using the Endo-Free Plasmid Maxi Kit (Qiagen, Hilden, Germany), and the concentration and purity were assessed by UV spectrophotometry and agarose gel electrophoresis.

    Marker:

    Article Title: New Clox Systems for Rapid and Efficient Gene Disruption in Candida albicans
    Article Snippet: The CaMET3 promoter region (1336 bp) was then PCR-amplified using Infusion cloning primers Clox-MET3p-F and Clox-MET3p-R ( ) and cloned between the Nhe I and Xma I sites in pLUCL2 in front of the cre gene using an In-Fusion HD cloning kit according to the manufacturer’s instructions (Clontech, California, USA) to generate the URA3-Clox cassette in the plasmid pLUMCL2. .. The URA3 marker in pLUMCL2 was then replaced with the NAT1 marker to generate the NAT1-Clox cassette in the plasmid pLNMCL.

    Variant Assay:

    Article Title: Differential gene expression profiling linked to tumor progression of splenic marginal zone lymphoma
    Article Snippet: The pTRE3G1 plasmid vector contains the following constructs: reverse tetracycline activator coding region derived from pCMV-TET3G (Takara Bio Inc., Shiga, Japan); an improved variant of the copepod Pontellina plumata green fluorescent protein (max GFP) coding region derived from pmaxGFP (Lonza, Basel, Switzerland); the GAPDH promoter from nucleotides –376 to + 183 relative to the transcription start site; a tetracycline-response element promoter derived from pTRE3G (Takara Bio); 5′ and 3′ miR-155 flunking region derived from pcDNA 6.2-GW/EmGFP-miR (Thermo Fisher Scientific); and a Luciferase shRNA coding region derived from pSingle-tTS-Anti-Luc (Takara Bio Inc.) for control shRNA. .. The DNA fragments were synthesized using the GeneArt Strings system (Thermo Fisher Scientific) and cloned into SalI-PciI restriction enzyme sites of pcDNA3.1 (Thermo Fisher Scientific) using the In-Fusion HD cloning kit (Takara Bio Inc.), according to the manufacturer’s instructions.

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    TaKaRa in fusion hd cloning kit
    In Fusion Hd Cloning Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 1032 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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