Structured Review

TaKaRa subcloning
Subcloning, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 890 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/subcloning/product/TaKaRa
Average 94 stars, based on 890 article reviews
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subcloning - by Bioz Stars, 2020-09
94/100 stars

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Amplification:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

Subcloning:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

Construct:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

Sequencing:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

Polymerase Chain Reaction:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

Generated:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.

other:

Article Title: Nonpolarized signaling reveals two distinct modes of 3D cell migration
Article Snippet: cDNA constructs and siRNAs The pECFP-Rac1 and -Cdc42 constructs ( ) were generated by subcloning the full-length sequence into the EcoRI–BamH1 sites of pECFP-C1 (Takara Bio Inc.). pYPet–p21-binding domain (PBD) was generated by subcloning the sequence encoding amino acids 65–150 of human Pak1 into the EcoRI–BamH1 sites of pYPet-C1. pYPet-C1 was constructed by subcloning the sequence corresponding to the fluorescent protein YPet into the AgeI–XhoI sites of pEGFP-C1 (Takara Bio Inc.).

Article Title: Chemotrap-1: an engineered soluble receptor that blocks chemokine-induced migration of metastatic cancer cells in vivo
Article Snippet: pGBT9-hCCL21 vector was generated by inserting a cDNA encoding the mature form of human CCL21 (GenBank Accession No: ; aa24-134) into the BamHI site of MATCHMAKER yeast two-hybrid (Y2H) system 2 vector pGBT9 (Clontech). pGBKT7-hCCL21 vector was generated by subcloning the BamHI CCL21 fragment from pGBT9-hCCL21 into the BamHI site of pGBKT7 (Clontech). pGADT7-hCCL21 and pGADT7-hCCL21ΔCOOH expression vectors were generated by subcloning the BamHI fragment (encoding CCL21 amino acids 24-134) or the BamHI-PstI fragment (encoding CCL21 aa24-102) from pGBKT7-CCL21 into pGADT7 expression vector (Clontech). pGADT7-mCCL21 and pGADT7-CCL27 expression vectors were generated by cloning the cDNAs encoding mature forms of mCCL21 (GenBank Acc: ; aa24-133) or hCCL27 (GenBank Acc: ; aa25-112) into pGADT7.

Article Title: Mediator subunit MED1 modulates intranuclear dynamics of the thyroid hormone receptor.
Article Snippet: The thyroid hormone receptors (TRs) mediate thyroid hormone (T < sub > 3 < /sub > )-dependent gene expression.

Article Title: Gene optimization leads to robust expression of human respiratory syncytial virus nucleoprotein and phosphoprotein in human cells and induction of humoral immunity in mice.
Article Snippet: Primers were designed with restriction endonuclease sites flanking the genes for subcloning into pShuttle (Clontech, Mountain View, USA).

Article Title: Differential Expression of KCNQ2 Splice Variants: Implications to M Current Function during Neuronal Development
Article Snippet: For coexpression with Q2S, an N-terminal fusion construct of Q2L with enhanced green fluorescent protein (EGFP) was generated by subcloning into pEGFPC1 vector (Clontech).

Article Title: Tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex
Article Snippet: Expression constructs pLVX-EF1α-IRES-Puro FLAG-tagged human Tspan15 was generated by subcloning the FLAG-Tspan15 sequence into the pLVX-EF1α-IRES-Puro lentiviral plasmid (Clontech, Mountain View, CA).

Article Title: Cell Cycle Regulated Phosphorylation of the Telomere-Associated Protein TIN2
Article Snippet: Plasmids pBabe-puro-Flag-TIN2WT , pBabe-puro-TIN2WT -HA, and pEGFP-N1-TIN2WT were generated by introducing, in frame, an N-terminal Flag or a C-terminal HA epitope-tag in the human TIN2 cDNA by PCR and subcloning the resultant cDNA into the EcoRI/HindIII sites of pBabe-puro . pBabe-puro-Flag, pMAL-c2x-Flag and pEGFP-N1 TIN2S295A , TIN2S330A , and the compound S295A/330A TIN2AA mutant were generated by introducing S295A, S330A, or S295A/S330A mutations into the aforementioned Flag-TIN2WT cDNA and subcloning the resultant cDNAs into the EcoRI/HindIII sites of the pBabe-puro vector, the pMAL-c2x vector (New England Lab), and the XhoI/HindIII sites of the pEGFP-N1 vector (Clontech). pBabe-puro-TIN2S295A -HA was generated by introducing the S295A into the aforementioned TIN2WT -HA cDNA and subcloning the resultant cDNA into the EcoRI/HindIII sites of the pBabe-puro vector. pQCXIP-Flag-TIN2WT was generated by subcloning the aforementioned Flag-TIN2WT cDNA into the NotI/AgeI sites of the pQCXIP vector (catalogue # 6315, Clontech). pcDNA-Flag-RSK2Y707A was a kind gift from Dr. Sally Kornbluth. pCMV-myc-TRF1 and pEYFP-C1-TPP1 were previously described. pSuper-retro-GFP-Neo-shTIN2-1 and -2 were generated by insert small hairpin RNA against TIN2 (5′-GGAGCACAUUCUUUGCCUG-3′ and 5′- CCAACCCAGGUCAUAUCUAAG-3′) into the BglII/HindIII sites of the pSuper-retro-GFP-Neo vector.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Ets Transcription Factors Control Epithelial Maturation and Transit and Crypt-Villus Morphogenesis in the Mammalian Intestine
Article Snippet: .. All constructs also contained a C-terminal nuclear localization sequence (NLS; PKKKRKV, from the SV40 large T antigen), added during the first PCR amplification step. pSG5-HA/Erm was generated by subcloning a full-length Erm cDNA, amplified from a mouse embryonic brain library by reverse transcriptase (RT)-PCR, into pTRE-HA (Clontech), and then subcloning of the HA-tagged insert into pSG5. pSG5-HA/Ets2 was generated by subcloning a full-length mouse Ets2 cDNA (generously provided by James Hagman, National Jewish Medical and Research Center, Denver, CO) into pCGN2-HA, and then subcloning the HA-tagged insert into pSG5. pSG5-HA/Elf3 was generated by subcloning HA-tagged full-length human Elf3 into pSG5. .. The reporter construct 8x(EBS)-TK-luciferase was generated by subcloning the 8xpal sequence (containing eight copies of the DNA-binding site GCAGGAAGCA from the rat stromelysin promoter) from 8xpal-pBLCAT into pA3 -TK-luciferase.