Plasmid Preparation:
Article Title: Injectable mRNA-engineered short fibers deliver temporally coupled mechano-genetic signals to reprogram Schwann cell cytoskeleton for nerve repair.
Article Snippet: To eliminate endotoxin contamination, plasmid templates were isolated using FastPure Enhanced EndoFree Plasmid Maxi Kit (DC212, Vazyme, Nanjing, China), guaranteeing endotoxin levels<0.1 EU/μg. .. The plasmid was linearized using the restriction enzyme XhoI (R0146M, New England Biolabs, Ipswich, MA) and transcribed into mRNA using T7 High Yield RNA Transcription Kit (E131, Novoprotein, Suzhou, China). ..
Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Article Title: Tryptophan oxidation-induced dynamics highlight why stable γS-crystallin variants aggregate.
Article Snippet: The gene fragment for His-tagged γS-W163S (Sup- 121 plementary Figure S1) was purchased from Twist Bioscience (South San Francisco, California). .. 122 Restriction enzymes NcoI and XhoI and Gibson assembly master mix were purchased from New 123 England Biolabs (Ipswich, Massachusetts) and used to assemble the pET28a(+) vector contain- 124 ing the gene for γS-W163S. .. Genes of interest within plasmid samples were verified by Sanger 125 sequencing using the T7 universal primer provided by GENEWIZ from Azenta (South Plainfield, 126 New Jersey).
Article Title: Characterization of novel Klebsiella pneumoniae phages isolated from hospital wastewater in Mumbai, India
Article Snippet: .. PCR products were digested by NdeI and XhoI ( dep1627_49, dep1627_50 , and depG66_49 ) or BamHI and XhoI ( depKBKp12_46 and dep1047_48 ) (New England BioLabs, USA) and cloned into expression vector pGD or pGD SUMO ( ) with an N-terminal histidine tag. .. Competent E. coli Turbo cells (New England BioLabs, USA) were transformed for plasmid propagation and isolation with subsequent Sanger sequencing.
Virus:Article Title: Metazoan lagging-strand DNA synthesis is distributive.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Anti-xPOLA1 This paper N/A Rabbit Anti-xPOLD1 This paper N/A Rabbit Anti-xGINS Terui et al.81 N/A Peroxidase AffiniPure Rabbit Anti-Mouse IgG (H + L) Jackson Immuno Research Labs Cat# 315-035-003; RRID:AB_2340061 Bacterial and virus strains DH5α NEB Cat# C2987H DH10EMBacY Geneva Biotech Cat# DH10EMBacY Rosetta (DE3) pLysS Novagen Cat# 70956-4 Chemicals, peptides, and recombinant proteins BamHI-HF NEB Cat# R3136S BstXI NEB Cat# R0113S HindIII-HF NEB Cat# R3104S NcoI-HF NEB Cat# R3193S NotI-HF NEB Cat# R3189S SalI-HF NEB Cat# R3138S SwaI NEB Cat# R0604S PI-SceI NEB Cat# R0696S PmeI NEB Cat# R0560S XhoI NEB Cat# R0146S Klenow Fragment (3′–5′ exo-) polymerase NEB Cat# M0212S Proteinase K NEB Cat# P8107S [α-32P] dCTP PerkinElmer Cat# BLU513A250UC biotin-dCTP Thermo Fisher Scientific Cat# 19518018 biotin-dGTP PerkinElmer Cat# NEL541001EA L-Cysteine hydrochloride monohydrate Fisher Scientific Cat# ICN10144601 Cycloheximide Sigma Aldrich Cat# 01810 Aprotinin Sigma Aldrich Cat# 11583794001 Leupeptin Sigma Aldrich Cat# 11529048001 Cytochalasin B from Drechslera dematioidea Sigma Aldrich Cat# C6762 Dithiothreitol (DTT) Bio-Rad Cat# 1610611 Nocodazole Sigma Aldrich Cat# M1404 Dimethyl sulfoxide (DMSO) Sigma Aldrich Cat# D8418 Adenosine 5′-triphosphate disodium salt hydrate (ATP) Sigma Aldrich Cat# A7699 Phosphocreatine disodium salt hydrate (PC) Sigma Aldrich Cat# P7956 Creatine Phosphokinase from rabbit muscle (CPK) Sigma Aldrich C3755 FEN1mkikGR Terui et al.81 N/A His6-Human Rhinovirus (HRV) 3C Protease Terui et al.81 N/A Alexa Fluor 647 C2 Maleimide Thermo Fisher Scientific Cat# A20347 Lumidyne 555 CoA Conjugate Lumidyne Technologies Custom Quote Lumidyne 655 CoA Conjugate Lumidyne Technologies Custom Quote (Continued on next page) ll OPEN ACCESS Cell 189, 1–12.e1–e10, December 24, 2026 e1 Short article .. REAGENT or RESOURCE SOURCE IDENTIFIER Ni-NTA Agarose Qiagen Cat# 30230 Imidazole Fisher Scientific Cat# 03196-500 Dynabeads Protein A Invitrogen Cat# 10002D rProtein A Sepharose Fast Flow GE Healthcare Cat# 171279 AminoLink Plus Coupling Resin Fisher Scientific Cat# PI20501 Cellfectin II Reagent Fisher Scientific Cat# 10-362-100 LB Broth Thermo Fisher Scientific Cat# BP9723 LB Agar Thermo Fisher Scientific Cat# BP9724 ESF 921 Insect Cell Culture Medium, Protein Free Expression Systems Cat# 96-001-01 Superdex 200 Increase 10/300 GL Cytiva Cat# 28990944 Mono Q 5/50 GL Cytiva Cat# 17516601 Mono S 5/50 GL Cytiva Cat# 17516801 Amicon Ultra-4-10K Merck Millipore Cat# UFC801024 Spectrum Spectra/Por 1 RC Dialysis Membrane Tubing 6–8 kD Spectrum Labs Cat# 132645 Spectrum Spectra/Por 4 RC Dialysis Membrane Tubing 12–14 kD Spectrum Labs Cat# 132700 Pierce Protease Inhibitor Tablets, EDTAfree ThermoFisher Scientific Cat# A32965 4%–15% Mini-PROTEAN TGX Precast Protein Gels Bio-Rad Cat# 4561086 Tris-Glycine-SDS Running Buffer Boston BioProducts Cat# BP-150 EZ-Run Prestained Rec Protein Ladder Fisher Scientific Cat# BP36031 Precision Plus Protein Dual Color Standard Bio-Rad Cat# 1610374EDU SuperSignal West Pico PLUS Chemiluminescent Substrate Fisher Scientific Cat# PI34577 Optiblot Blue InstantBlue Coomassie Protein Stain Abcam Cat# ab119211 PVDF membranes VWR Cat# 10061-492 Transfer Buffer Fisher Scientific Cat# NC9297917 Phosphate Buffered Saline (10x, pH 7.4) Fisher Scientific Cat# NC9140736 CHORULON Hormone Merck Cat# 133754 75 × 25 × 1 mm Glass Slides VWR Cat# 48300-026 High Precision Glass CoverSlip, No 1.5, 24 × 60 mm Bioscience Tools Cat# CSHP-No1.5-24x60 Double-sided tape Grace Bio-Labs SKU 62001 mPEG-Succinimidyl Valerate, MW 5,000 Laysan Bio Cat# MPEG-SVA-5000-1g Biotin-PEG-SVA, MW 5,000 Laysan Bio Cat# Biotin-PEG-SVA-5000-100MG Streptavidin from Streptomyces avidinii Sigma Aldrich Cat# S4762-5MG Streptavidin, Alexa Fluor 647 conjugate Thermo Fisher Scientific Cat# S21374 Aphidicolin from Nigrospora sphaerica Sigma-Aldrich Cat# A0781-5MG Critical commercial assays Q5 High-Fidelity 2X Master Mix NEB Cat# M0492S NEBuilder HiFi DNA Assembly Master Kit NEB Cat# E2621S ZR BAC DNA Miniprep kit Zymo Research Cat# D4049 Deposited data Raw data from this paper Stanford Digital Repository https://doi.org/10.25740/sq803bx0564 Experimental models: Cell lines Sf9 insect cells Expression Systems Cat# 94-001S (Continued on next page) ll OPEN ACCESS e2 Cell 189, 1–12.e1–e10, December 24, 2026 Short article
Recombinant:Article Title: Metazoan lagging-strand DNA synthesis is distributive.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Anti-xPOLA1 This paper N/A Rabbit Anti-xPOLD1 This paper N/A Rabbit Anti-xGINS Terui et al.81 N/A Peroxidase AffiniPure Rabbit Anti-Mouse IgG (H + L) Jackson Immuno Research Labs Cat# 315-035-003; RRID:AB_2340061 Bacterial and virus strains DH5α NEB Cat# C2987H DH10EMBacY Geneva Biotech Cat# DH10EMBacY Rosetta (DE3) pLysS Novagen Cat# 70956-4 Chemicals, peptides, and recombinant proteins BamHI-HF NEB Cat# R3136S BstXI NEB Cat# R0113S HindIII-HF NEB Cat# R3104S NcoI-HF NEB Cat# R3193S NotI-HF NEB Cat# R3189S SalI-HF NEB Cat# R3138S SwaI NEB Cat# R0604S PI-SceI NEB Cat# R0696S PmeI NEB Cat# R0560S XhoI NEB Cat# R0146S Klenow Fragment (3′–5′ exo-) polymerase NEB Cat# M0212S Proteinase K NEB Cat# P8107S [α-32P] dCTP PerkinElmer Cat# BLU513A250UC biotin-dCTP Thermo Fisher Scientific Cat# 19518018 biotin-dGTP PerkinElmer Cat# NEL541001EA L-Cysteine hydrochloride monohydrate Fisher Scientific Cat# ICN10144601 Cycloheximide Sigma Aldrich Cat# 01810 Aprotinin Sigma Aldrich Cat# 11583794001 Leupeptin Sigma Aldrich Cat# 11529048001 Cytochalasin B from Drechslera dematioidea Sigma Aldrich Cat# C6762 Dithiothreitol (DTT) Bio-Rad Cat# 1610611 Nocodazole Sigma Aldrich Cat# M1404 Dimethyl sulfoxide (DMSO) Sigma Aldrich Cat# D8418 Adenosine 5′-triphosphate disodium salt hydrate (ATP) Sigma Aldrich Cat# A7699 Phosphocreatine disodium salt hydrate (PC) Sigma Aldrich Cat# P7956 Creatine Phosphokinase from rabbit muscle (CPK) Sigma Aldrich C3755 FEN1mkikGR Terui et al.81 N/A His6-Human Rhinovirus (HRV) 3C Protease Terui et al.81 N/A Alexa Fluor 647 C2 Maleimide Thermo Fisher Scientific Cat# A20347 Lumidyne 555 CoA Conjugate Lumidyne Technologies Custom Quote Lumidyne 655 CoA Conjugate Lumidyne Technologies Custom Quote (Continued on next page) ll OPEN ACCESS Cell 189, 1–12.e1–e10, December 24, 2026 e1 Short article .. REAGENT or RESOURCE SOURCE IDENTIFIER Ni-NTA Agarose Qiagen Cat# 30230 Imidazole Fisher Scientific Cat# 03196-500 Dynabeads Protein A Invitrogen Cat# 10002D rProtein A Sepharose Fast Flow GE Healthcare Cat# 171279 AminoLink Plus Coupling Resin Fisher Scientific Cat# PI20501 Cellfectin II Reagent Fisher Scientific Cat# 10-362-100 LB Broth Thermo Fisher Scientific Cat# BP9723 LB Agar Thermo Fisher Scientific Cat# BP9724 ESF 921 Insect Cell Culture Medium, Protein Free Expression Systems Cat# 96-001-01 Superdex 200 Increase 10/300 GL Cytiva Cat# 28990944 Mono Q 5/50 GL Cytiva Cat# 17516601 Mono S 5/50 GL Cytiva Cat# 17516801 Amicon Ultra-4-10K Merck Millipore Cat# UFC801024 Spectrum Spectra/Por 1 RC Dialysis Membrane Tubing 6–8 kD Spectrum Labs Cat# 132645 Spectrum Spectra/Por 4 RC Dialysis Membrane Tubing 12–14 kD Spectrum Labs Cat# 132700 Pierce Protease Inhibitor Tablets, EDTAfree ThermoFisher Scientific Cat# A32965 4%–15% Mini-PROTEAN TGX Precast Protein Gels Bio-Rad Cat# 4561086 Tris-Glycine-SDS Running Buffer Boston BioProducts Cat# BP-150 EZ-Run Prestained Rec Protein Ladder Fisher Scientific Cat# BP36031 Precision Plus Protein Dual Color Standard Bio-Rad Cat# 1610374EDU SuperSignal West Pico PLUS Chemiluminescent Substrate Fisher Scientific Cat# PI34577 Optiblot Blue InstantBlue Coomassie Protein Stain Abcam Cat# ab119211 PVDF membranes VWR Cat# 10061-492 Transfer Buffer Fisher Scientific Cat# NC9297917 Phosphate Buffered Saline (10x, pH 7.4) Fisher Scientific Cat# NC9140736 CHORULON Hormone Merck Cat# 133754 75 × 25 × 1 mm Glass Slides VWR Cat# 48300-026 High Precision Glass CoverSlip, No 1.5, 24 × 60 mm Bioscience Tools Cat# CSHP-No1.5-24x60 Double-sided tape Grace Bio-Labs SKU 62001 mPEG-Succinimidyl Valerate, MW 5,000 Laysan Bio Cat# MPEG-SVA-5000-1g Biotin-PEG-SVA, MW 5,000 Laysan Bio Cat# Biotin-PEG-SVA-5000-100MG Streptavidin from Streptomyces avidinii Sigma Aldrich Cat# S4762-5MG Streptavidin, Alexa Fluor 647 conjugate Thermo Fisher Scientific Cat# S21374 Aphidicolin from Nigrospora sphaerica Sigma-Aldrich Cat# A0781-5MG Critical commercial assays Q5 High-Fidelity 2X Master Mix NEB Cat# M0492S NEBuilder HiFi DNA Assembly Master Kit NEB Cat# E2621S ZR BAC DNA Miniprep kit Zymo Research Cat# D4049 Deposited data Raw data from this paper Stanford Digital Repository https://doi.org/10.25740/sq803bx0564 Experimental models: Cell lines Sf9 insect cells Expression Systems Cat# 94-001S (Continued on next page) ll OPEN ACCESS e2 Cell 189, 1–12.e1–e10, December 24, 2026 Short article
Hemagglutination Assay:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Expressing:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Article Title: Characterization of novel Klebsiella pneumoniae phages isolated from hospital wastewater in Mumbai, India
Article Snippet: .. PCR products were digested by NdeI and XhoI ( dep1627_49, dep1627_50 , and depG66_49 ) or BamHI and XhoI ( depKBKp12_46 and dep1047_48 ) (New England BioLabs, USA) and cloned into expression vector pGD or pGD SUMO ( ) with an N-terminal histidine tag. .. Competent E. coli Turbo cells (New England BioLabs, USA) were transformed for plasmid propagation and isolation with subsequent Sanger sequencing.
Cloning:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Bioprocessing:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Software:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Incubator:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Spectrophotometry:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Gel Electrophoresis:Article Title: Endogenous Protein Tagging in Postmitotic Neurons via Lentiviral and Adeno‐associated Virus Delivery
Article Snippet: .. pLentiSIV hSyn HA‐NLS‐SpCas9‐NLS WPRE (SIV‐based lentiviral backbone allowing the Lentiviral expression of SpCas9; Addgene plasmid # 236242 T4 DNA Ligase and Reaction Buffer (NEB, cat. no. M0202) T4 Polynucleotide Kinase (T4 PNK; NEB, cat. no. M0201) Oligonucleotides for sgRNA cloning (IDT; see protocol steps for sequences) BbsI‐HF restriction enzyme, (NEB, cat. no. R3539), HindIII‐ HF (NEB, cat. no. R3104), XhoI restriction enzymes (NEB, cat. no. R0146), and rCutSmart Buffer pAAV KI Cloning Vector: pAAV KI Cloning Vector_hSyn mTagBFP2‐CAAX2 (AAV backbone with U6 promoter, BbsI sgRNA cloning sites, gRNA scaffold, HindIII/XhoI donor insertion sites, and hSyn‐mTagBFP2‐CAAX2 reporter); Addgene plasmid #236245 Competent E. coli : DH5α (NEB, C2988J) or Stbl3 for viral plasmids (One Shot Stbl3 Chemically Competent E. coli , Invitrogen, cat. no. C737303) SOC medium (Sigma‐Aldrich, cat. no. S1797) LB agar plates with 100 μg/mL ampicillin (as described in Repository and Christensen, ) In‐Fusion HD Cloning Kit (Takara Bio, cat. no. 639648) or equivalent Gibson assembly kit Plasmid miniprep and maxiprep kits Annealing buffer: 10 mM Tris·HCl (pH 7.5), 50 mM NaCl, and 1 mM EDTA NEBNext Ultra II Q5 Master Mix (NEB, cat. no. M0544) Benchling molecular biology software ( https://benchling.com ) 37°C bacterial incubator with shaker Thermocycler UV spectrophotometer (NanoDrop or equivalent) Gel electrophoresis apparatus ..
Gibson Assembly:Article Title: Tryptophan oxidation-induced dynamics highlight why stable γS-crystallin variants aggregate.
Article Snippet: The gene fragment for His-tagged γS-W163S (Sup- 121 plementary Figure S1) was purchased from Twist Bioscience (South San Francisco, California). .. 122 Restriction enzymes NcoI and XhoI and Gibson assembly master mix were purchased from New 123 England Biolabs (Ipswich, Massachusetts) and used to assemble the pET28a(+) vector contain- 124 ing the gene for γS-W163S. .. Genes of interest within plasmid samples were verified by Sanger 125 sequencing using the T7 universal primer provided by GENEWIZ from Azenta (South Plainfield, 126 New Jersey).
Polymerase Chain Reaction:Article Title: Characterization of novel Klebsiella pneumoniae phages isolated from hospital wastewater in Mumbai, India
Article Snippet: .. PCR products were digested by NdeI and XhoI ( dep1627_49, dep1627_50 , and depG66_49 ) or BamHI and XhoI ( depKBKp12_46 and dep1047_48 ) (New England BioLabs, USA) and cloned into expression vector pGD or pGD SUMO ( ) with an N-terminal histidine tag. .. Competent E. coli Turbo cells (New England BioLabs, USA) were transformed for plasmid propagation and isolation with subsequent Sanger sequencing.
Clone Assay:Article Title: Characterization of novel Klebsiella pneumoniae phages isolated from hospital wastewater in Mumbai, India
Article Snippet: .. PCR products were digested by NdeI and XhoI ( dep1627_49, dep1627_50 , and depG66_49 ) or BamHI and XhoI ( depKBKp12_46 and dep1047_48 ) (New England BioLabs, USA) and cloned into expression vector pGD or pGD SUMO ( ) with an N-terminal histidine tag. .. Competent E. coli Turbo cells (New England BioLabs, USA) were transformed for plasmid propagation and isolation with subsequent Sanger sequencing.
|
New England Biolabs is a verified supplier 
New England Biolabs manufactures this product 