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Citations for New England Biolabs :
101 - 150 of 3408 citations for pLenti CMV TO Puro DEST 670 1 since 2020
Citations are collected from bioRxiv only, the total number of publications could be much larger.
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bioRxiv - Synthetic Biology 2024Quote: Transcription templates for expressing gRNA variants and SARS-CoV-2 or CMV input RNA fragments were generated by PCR (Phusion high-fidelity PCR kit, NEB #E0553) of the gBlock or Ultramer template that included a T7 promoter and the gRNA or input RNA coding sequence ...
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bioRxiv - Microbiology 2022Quote: ... The pLVX-StrepII-ORF-IRES-Puro plasmids were then cloned using the NEBuilder HiFi DNA Assembly Cloning Kit (NEB) from the pTriEX-6 plasmids ...
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bioRxiv - Genomics 2021Quote: ... and then inserted into the BsmBI-digested lentiCRISPR-v2-Blast-Puro vector by Gibson assembly (New England Biolabs, E2621L). The assembled products were transformed into NEB 5-alpha electrocompetent E ...
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bioRxiv - Microbiology 2023Quote: ... and corresponding gene blocks were synthesized (IDT) and assembled with the pcDNA3.1-puro backbone by HiFi DNA Assembly (NEB). All amino acid sequences were codon-optimized for expression in human cells ...
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bioRxiv - Cancer Biology 2024Quote: ... Specific mutations in human PSEN1 were obtained by Q5 site-directed mutagenesis of the pMSCV-puro vector containing wild type human PSEN1.26 We designed primers for Q5 site-directed mutagenesis (New England Biolabs) with NEBaseChanger.neb.com and performed site-directed mutagenesis according to the manufacturer’s protocol ...
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bioRxiv - Molecular Biology 2023Quote: ... and inserted into the pKH011-puro backbone by simultaneous digestion of the backbone with BbsI-HF (NEB, Cat# R3539S) and ligation with T4 DNA ligase (NEB ...
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bioRxiv - Immunology 2024Quote: ... the double-stranded DNA sgRNA were individually cloned into the pLKO1.5-puro-GFP vector using T4 DNA ligase (New England Biolabs), and the plasmids were transformed into chemically competent Stbl3 E.coli (Thermo Fisher) ...
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bioRxiv - Cell Biology 2022Quote: ... mTagBFP2 and Cas9-Avidin provided by Max Wilson into an expression vector bearing a CMV promoter and flanking PiggyBac transposase-compatible inverted terminal repeats using Gibson Assembly (New England BioLabs Inc., E2611L) according to supplier instructions ...
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bioRxiv - Synthetic Biology 2024Quote: ... The concentrated fragments (NheI-CMV-fp-MfeI) were subsequently isolated using the Monarch Gel DNA Extraction Kit (New England Biolabs, Cat# T1020L). The concentration of each purified probe was measured by a Qubit Flex fluorometer (Invitrogen by Thermo Fisher Scientific ...
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bioRxiv - Synthetic Biology 2024Quote: ... DNA fragments (NheI-CMV-fp-tPT2A-fp-MfeI) containing MuSIC barcodes were isolated by the Monarch Gel DNA Extraction Kit (New England Biolabs, Cat# T1020L) and subject to end-prep ...
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bioRxiv - Cell Biology 2020Quote: ... photoactivatable-mCherry was PCR-amplified from the plasmid N-PA-mCh and assembled into the retroviral vector pBABE-puro using HiFi DNA Assembly (E5520S, NEB). To create the stable U2OS-dCas9-PA-mCh/GFP-Parkin and U2OS-dCas9-PA-mCh/TFEB-GFP cell lines ...
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bioRxiv - Genomics 2020Quote: The lentiviral vector for expressing TIR1 (Lentiv4-EFsp-Puro-2A-TIR1-9Myc) was constructed using PCR and Gibson Assembly Cloning kit (New England Biolabs). The backbone was modified from lentiCRISPR v259 (a gift from Feng Zhang ...
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bioRxiv - Genomics 2020Quote: ... The oligo-pool amplicons were assembled into the linearized Lenti-gRNA-Puro plasmid using NEBuilder HiFi DNA Assembly Master Mix (NEB) for 1 h at 50 °C ...
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bioRxiv - Cell Biology 2020Quote: ... CASP1 and CASP4 CDS were amplified from the obtained library and cloned into the pMSCV-puro vectors The caspase-4 catalytically dead C258A pMSCV-puro vector was generated from the wild-type pMSCV-puro-CASP4 through site-directed mutagenesis by PCR using the Q5 Site-Directed Mutagenesis Kit (New England Biolabs). The CASP4 and GSDMD-targeting lentiviral vectors (pGIPZ ...
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bioRxiv - Genetics 2020Quote: ... including protospacer sequences were amplified by PCR to create homology arms and cloned into lentiCRISPR v2 FE Puro backbone through NEBuilder HiFi DNA assembly (NEB). Protospacer sequences used are listed below:
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bioRxiv - Immunology 2021Quote: ... and Slc2a8 knockout cell line were prepared by digesting lentiGuide-puro constructs with already cloned gRNAs sequences with BsiWI-HF and MluI-HF (R0553 and R3198, NEB) to exclude the puromycin N-acetyltransferase gene ...
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bioRxiv - Genetics 2020Quote: ... The ‘2A-Puro-WPRE’ sequence was then removed from the LCv2 plasmid via restriction digestion with PmeI (NEB, Cat# R0560S) and BamHI (NEB ...
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bioRxiv - Microbiology 2021Quote: ... Insertion of hACE2 into pLVX-IRES-puro was conducted by doubl e digestion of XhoI and XbaI (Fermantas) and ligation of T4 ligase (NEB) according to manufact urer’s instructions ...
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bioRxiv - Cell Biology 2023Quote: HPV-E6/E7 genomic DNA was subcloned into a pLVX-Tight-Puro backbone (a gift from Genmedic, China) downstream of the Tet operator by the restriction enzyme cutting site EcoRI (NEB) and BamHI (NEB) ...
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bioRxiv - Cell Biology 2023Quote: ... the repair vector GW209_pCRIS-PITChv2-C-dTAG-Puro (BRD4) (2 μg) was digested with MluI-HF (New England Biolabs; #R3198) in Cutsmart Buffer for 1 hour ...
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bioRxiv - Cancer Biology 2024Quote: ... Base editing sgRNA directed against the SF3B1 K700 locus was designed using BE-Designer28 and cloned into pLKO.5 Puro-2A-GFP between BsmBI (New England Biolabs) cut sites ...
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bioRxiv - Cancer Biology 2024Quote: ... the hU6-pegRNA2-polyT cassette was amplified from the pU6-pegRNA-gg-acceptor backbone and ligated into ngRNA1 pLKO.5 Puro-2A-RFP between EcoRI and XhoI (New England Biolabs) to create pLKO.5-K700E-ng1+pg2-Puro-2A-RFP ...
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bioRxiv - Cell Biology 2023Quote: ... Cas9 plasmids targeting sequences BLTP2 open reading frame were prepared by digesting pSpCas9(BB)-2A-Puro plasmid with BbsI (R3539S, NEB) and inserting duplexed primers caccGAGCCAAACACGGTTCCGGA and aaacTCCGGAACCGTGTTTGGCTC or caccGTGCAAGATCTGACGCCGAC and aaacGTCGGCGTCAGATCTTGCAC ...
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bioRxiv - Molecular Biology 2024Quote: ... complementary synthesized oligonucleotides (Integrated DNA technologies) were annealed and ligated into a lentiGuide- Puro plasmid after BsmBI digestion (New England Biolabs). For the expression of MS2-sgRNAs ...
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bioRxiv - Genetics 2024Quote: ... The oligo pool encoding guides (Supplementary table T10) were synthesized by Twist Bio and inserted into addgene Plasmid #52963 lentiGuide-Puro digested with Esp3I enzyme (NEB). To evaluate the effects of CRISPR interference cells were transduced with sgRNA lentiviruses ...
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bioRxiv - Developmental Biology 2024Quote: ... the 6xHis-EGFP-HRV3C and Trus coding sequences (BDGP: RE69372) were assembled together on the pMT-puro vector using NEBuilder HiFi DNA Assembly (New England Biolabs). EGFP-tagged Trus expression induced with da-GAL4 in vivo completely rescued the trus mutant (trus4-15/Dftrus ...
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bioRxiv - Cancer Biology 2022Quote: ... cDNA of EWSR1 and MN1 were obtained from the DKFZ cDNA clone repository and assembled with an HA-tag at the N-terminus and SSX at the C-terminus into a MSCV-PGK-Puro backbone in a single step using NEBuilder HiFi DNA Assembly (NEB, #E2621).
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bioRxiv - Evolutionary Biology 2020Quote: ... The ZEB2 ORF was amplified with primers ZEB2_IndOE_F & _R and tagged with GFP-Flag by Gibson assembly into the pT2-CAG-MCS-2A-Puro plasmid linearized by restriction digestion with EcoRI (NEB, R3101S) and AgeI (NEB ...
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bioRxiv - Developmental Biology 2021Quote: ... A D10A mutation was introduced by site-directed mutagenesis of the original Puro-Cas9 donor with the Q5 mutagenesis kit (New England Biolabs, E0554S) to generate the Cas9n ...
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bioRxiv - Immunology 2021Quote: ... and after hybridization were cloned into Esp3I-digested lentiGuide-puro using the NEBuilder® HiFi DNA Assembly Master mix (E2621, NEB). Successful constructs were verified in plasmid from isolated colonies by sequencing with the LKO.1 5’ primer ...
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bioRxiv - Molecular Biology 2023Quote: Plasmid pMT-BRWD3-V5 was created by inserting the BRWD3 CDS sequence into the pMT-puro vector (addgene #17923) using the Gibson assembly method provided by NEB (# E5510S). For the pUbi63E-Flag-Ubiquitin plasmid ...
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bioRxiv - Biophysics 2024Quote: ... and the amplicon was inserted downstream of the CMV promoter sequence via Gibson assembly with NEBuilder® HiFi DNA Assembly Master Mix (NEB, Ipswich, MA, USA) into the pEGFP-N2 backbone ...
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bioRxiv - Cell Biology 2022Quote: pLV-PKR-GFP was generated from pPKR-GFP and pLV-EF1-Cre-PGK-Puro using NEBuilder® HiFi DNA Assembly (NEB #E5520) and the following primers ...
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MIIP downregulation promotes colorectal cancer progression via inducing adjacent adipocytes browningbioRxiv - Cancer Biology 2023Quote: ... Miip CDS was amplified by PCR and cloned into LV17 (EF-1α-Luciferase-puro) shuttle vector (GenePharma, Shanghai, China) with restriction enzymes Not I and Bam HI (New England Biolabs, Ipswich, MA). For mouse Miip specific knock-down ...
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bioRxiv - Genomics 2023Quote: ... The double-stranded lenti_epegRNA oligos were then assembled with the linearized Lenti-epeg-Puro-P2A-EGFP backbone using NEBuilder® HiFi DNA Assembly Master Mix (New England Biolabs, Cat. No. E2621S) using the standard protocol ...
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bioRxiv - Genomics 2022Quote: ... 2016a)] or pU6-sgRNA EF1Alpha-puro-T2A-mCherry [a gift from Gregory Ow and Eric Collisson, UCSF, (Jost et al., 2020)] digested with BstXI (NEB or Thermo Fisher Scientific) and BlpI (NEB ...
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bioRxiv - Genomics 2022Quote: ... 1 μl of Exonuclease 1 (NEB, M0293S) was added to each PCR product and incubated at 37C for 15 min to digest any single-stranded product ...
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bioRxiv - Molecular Biology 2024Quote: ... in 1× NEB buffer 1 (NEB, B7001S) for 15 min at 37°C ...
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bioRxiv - Biochemistry 2023Quote: ... 1% cholorophorm was added together with 10 μg mL-1 DNase 1 (NEB) and 1 μg mL-1 RNase A ...
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bioRxiv - Molecular Biology 2024Quote: ... The four PCR products were then assembled in a 1:1:1:1 ratio using HiFi assembly (NEB, E5520S) according to the manufacturer’s instructions to create the pLenti-STARR vector (pAS5018) ...
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bioRxiv - Microbiology 2022Quote: The recombinant constructs (pETDuet-1-TaPHB-1 and pETDuet-1-TaPHB-1-RUVBL1 were separately transformed into E. coli Lemo21(DE3) (NEB) chemical competent cells ...
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bioRxiv - Molecular Biology 2023Quote: ... Unincorporated ddRTP molecules were inactivated by adding 1 µL of Buffer 2 (80 mM MgCl2) and 1 µL of 1:1 rSAP (NEB):50% glycerol ...
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bioRxiv - Developmental Biology 2024Quote: ... 1 μL T4 RNA Ligase 1 (NEB, M0437M), 1 μL SuperAseIn (Invitrogen AM2694) ...
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bioRxiv - Molecular Biology 2023Quote: ... 1 mM MgCl2) supplemented with 1 mM ATP and 1 U/μL RNase Inhibitor (NEB). Reaction was incubated for 1 hour at 37°C ...
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bioRxiv - Molecular Biology 2024Quote: ... Plasmids were diluted 1:1 and 1:5 respectively in puc19 vector (New England Biolabs). HaloTag expression was confirmed using HaloTag ligand dyes HTL-AF488 ...
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bioRxiv - Molecular Biology 2020Quote: ... 1 unit µL-1 EcoRI-HF (New England Biolabs)) ...
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bioRxiv - Biophysics 2020Quote: ... 1 U μL−1 murine RNase inhibitor (NEB, M0314L), 600 ng mRNA and 50 μM PF846 in 1% DMSO ...
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bioRxiv - Genomics 2021Quote: ... 1 unit/μl RNA ligase 1 (New England Biolabs), 1× RNA ligase buffer ...
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bioRxiv - Microbiology 2023Quote: ... 1 μl 40 U/μl−1 RNase Inhibitor (NEB), and 1 μl T4 RNA ligase 2 (truncated K227Q ...
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bioRxiv - Immunology 2023Quote: ... supplemented with 1 U μl−1 RNase inhibitor (NEB), or bulk sorted into buffer RLT Plus (Qiagen ...