sapi  (New England Biolabs)


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  • 94
    Name:
    SapI
    Description:
    SapI 1 250 units
    Catalog Number:
    R0569L
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    1 250 units
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    Structured Review

    New England Biolabs sapi
    SapI
    SapI 1 250 units
    https://www.bioz.com/result/sapi/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Development of the SapI/AarI Incision Mediated Plasmid Editing Method"

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method

    Journal: Journal of molecular biology

    doi: 10.1016/j.jmb.2018.03.030

    Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.
    Figure Legend Snippet: Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.

    Techniques Used: Sequencing, Mutagenesis

    2) Product Images from "A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology"

    Article Title: A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2013.00339

    Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.
    Figure Legend Snippet: Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.

    Techniques Used: Plasmid Preparation, Clone Assay, Produced

    3) Product Images from "CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo"

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo

    Journal: Current protocols in molecular biology

    doi: 10.1002/cpmb.46

    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.
    Figure Legend Snippet: Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Techniques Used: Plasmid Preparation, Clone Assay, Construct

    Related Articles

    Clone Assay:

    Article Title: Antibody Responses toward the Major Antigenic Sites of Influenza B Virus Hemagglutinin in Mice, Ferrets, and Humans
    Article Snippet: .. The HA segments were subsequently cloned into a SapI (New England BioLabs)-digested pDZ ambisense vector ( ) through In-Fusion HD cloning (Clontech). ..

    Article Title: Distinct subnetworks of the thalamic reticular nucleus
    Article Snippet: .. The sgRNA library backbone was digested with SapI endonuclease (New England Biolabs, #R0569), and annealed sgRNA inserts were cloned into the backbone by Golden Gate assembly. ..

    Plasmid Preparation:

    Article Title: Antibody Responses toward the Major Antigenic Sites of Influenza B Virus Hemagglutinin in Mice, Ferrets, and Humans
    Article Snippet: .. The HA segments were subsequently cloned into a SapI (New England BioLabs)-digested pDZ ambisense vector ( ) through In-Fusion HD cloning (Clontech). ..

    Article Title: Production of single- and multiple-gene-modified mice via maternal SpCas9-based gene editing
    Article Snippet: .. Linearize the pCGsapI vector by SapI digestion (NEB). ..

    Article Title: Plug and Play Protein Modification using Homology-independent Universal Genome Engineering
    Article Snippet: .. In brief, 10ng (~2fmol) of the GS-gRNA backbone plasmid was digested with SapI enzyme (NEB R0569, 1uL) and simultaneously ligated with 50fmol of annealed 23-24mer (including SapI sticky ends) GS-gRNA oligos with T4 DNA ligase (NEB M0202, 1uL) in a 10uL reaction, by 10 repeats of thermocycling between 37°C (5 min) and 21°C (5 min). ..

    In Vitro:

    Article Title: Coxsackievirus B3 Responds to Polyamine Depletion via Enhancement of 2A and 3C Protease Activity
    Article Snippet: .. Briefly, CVB3 infectious clone [ ] was linearized with SapI (New England Biolabs [NEB], Ipswich, MA, USA) and used to generate RNA in vitro (NEB). ..

    Polymerase Chain Reaction:

    Article Title: Model-driven engineering of N-linked glycosylation in Chinese Hamster Ovary cells
    Article Snippet: .. The fragments were assembled using one-pot SapI restriction digestion-ligation reaction from multiple PCR fragments (using 10 U SapI (New England Biolabs, #R0569) and 5 U T4 ligase (Promega, # M1804). ..

    Sequencing:

    Article Title: Predictable and precise template-free CRISPR editing of pathogenic variants
    Article Snippet: .. Library integrity was verified by restriction digest with SapI (New England Biolabs) for 1 hour at 37°C, and sequence diversity was validated by high-throughput sequencing (HTS) as described below. ..

    High Throughput Screening Assay:

    Article Title: Predictable and precise template-free CRISPR editing of pathogenic variants
    Article Snippet: .. Library integrity was verified by restriction digest with SapI (New England Biolabs) for 1 hour at 37°C, and sequence diversity was validated by high-throughput sequencing (HTS) as described below. ..

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    New England Biolabs sapi restriction sites
    SapTrap assembly of MosSCI targeting vectors using the five-cassette system. A. Schematic of <t>pXF87</t> and the donor cassettes following <t>SapI</t> digestion. The dotted lines indicate the overhangs that anneal during ligation. B. Summary of available promoter, gene tag and 3’UTR donor cassette plasmids for the five-cassette system.
    Sapi Restriction Sites, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sapi restriction sites/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi restriction sites - by Bioz Stars, 2021-09
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    94
    New England Biolabs sapi enzyme
    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed <t>oligos</t> corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and <t>SapI</t> sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.
    Sapi Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sapi enzyme/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi enzyme - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    94
    New England Biolabs sapi restriction enzyme
    Ube3b knockout in mice results in growth retardation, failure to thrive and an overall smaller brain. Related to the entire paper. (A) Gene targeting strategy of murine Ube3b . The domain structure of Ube3b, wild type, and targeted Ube3b alleles are depicted. Exons, loxP sites, and FLP-recombinase target sites are symbolized as black rectangles, red triangles, and green rectangles, respectively. Exon 7 of Ube3b is flanked by two loxP sites. Mice positive for the recombined allele were crossed with FLP deleter animals to remove the lacZ/neomycin selection cassette, and further crossed to Cre-driver lines, leading to removal of exon 7 conditionally or conventionally. (B, C, D) Validation of Ube3b gene targeting. +/+ wild type, rec/+ indicates heterozygous recombined mutant. (B) The result of long-range PCR using primers depicted as blue arrows in A. Band of the predicted size was observed only using genomic <t>DNA</t> purified from rec/+ mutant as a template. (C) The result of multiplex PCR using primers depicted as pink arrows in A. PCR using genomic DNA from targeted ES cells yields two bands (left lane), and PCR using wild type ES cells genomic DNA as a template results in one band. (D) Southern blotting analysis of genomic DNA purified from targeted ES cells. Genomic DNA isolated from control and ES cells carrying ‘Recombined’ allele of Ube3b was digested with <t>SapI</t> enzyme. The probe is indicated as a green line in A. The band at 8.0 kb represents the wild type allele, and the band at 6.6 kb represents the mutant allele. (E) Gross morphology of Ube3b +/+ (left) and Ube3b -/- (right) mice at 20 days post birth. (F) Brains of Ube3b +/+ (left) and Ube3b -/- (right) mice isolated 20 days after birth. (G) Results of Western blotting using P7 brain lysates from Ube3b +/+ (left), Ube3b +/- (middle), and Ube3b -/- (right) animals with an antibody raised against HECT domain of Ube3b. MEM Code staining (lower panel) shows comparable amounts of total protein in all lanes.
    Sapi Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sapi restriction enzyme/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi restriction enzyme - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    SapTrap assembly of MosSCI targeting vectors using the five-cassette system. A. Schematic of pXF87 and the donor cassettes following SapI digestion. The dotted lines indicate the overhangs that anneal during ligation. B. Summary of available promoter, gene tag and 3’UTR donor cassette plasmids for the five-cassette system.

    Journal: bioRxiv

    Article Title: SapTrap assembly of C. elegans MosSCI transgene vectors

    doi: 10.1101/805507

    Figure Lengend Snippet: SapTrap assembly of MosSCI targeting vectors using the five-cassette system. A. Schematic of pXF87 and the donor cassettes following SapI digestion. The dotted lines indicate the overhangs that anneal during ligation. B. Summary of available promoter, gene tag and 3’UTR donor cassette plasmids for the five-cassette system.

    Article Snippet: Cloning To generate the expression vector pXF87, the two SapI restriction sites in pCFJ350 (Frøkjær-Jensen et al., 2012) were mutated using Q5 Site-Directed Mutagenesis (New England Biolabs) with the oligo pairs XF30F/XF30R and XF31F/XF31R.

    Techniques: Ligation

    SapTrap assembly of MosSCI targeting vectors using the four-cassette system. A. The MosSCI targeting vector pXF87 was derived from pCFJ350 by mutating two SapI restriction sites (indicated by arrowheads in the “Left” (L) and “Right” (R) homology arms) and introducing two SapI sites (blue text) between the XhoI and SpeI sites (green text). SapI cleavage sites are in red text. The SapI recognition sites are oriented such that upon digestion they are removed from the vector backbone. The cbr- unc-119 gene is used as a positive selection marker to facilitate the identification of transgenic animals. B. Design of the donor cassette vectors used for the 4- cassette cloning strategy. C. The curved dotted lines indicate the overhangs that anneal during the ligation reaction. D. Overview of the assembly protocol. For a detailed protocol, see the Materials and Methods section. E. Summary of available promoter, gene tag and 3’UTR donor cassette plasmids.

    Journal: bioRxiv

    Article Title: SapTrap assembly of C. elegans MosSCI transgene vectors

    doi: 10.1101/805507

    Figure Lengend Snippet: SapTrap assembly of MosSCI targeting vectors using the four-cassette system. A. The MosSCI targeting vector pXF87 was derived from pCFJ350 by mutating two SapI restriction sites (indicated by arrowheads in the “Left” (L) and “Right” (R) homology arms) and introducing two SapI sites (blue text) between the XhoI and SpeI sites (green text). SapI cleavage sites are in red text. The SapI recognition sites are oriented such that upon digestion they are removed from the vector backbone. The cbr- unc-119 gene is used as a positive selection marker to facilitate the identification of transgenic animals. B. Design of the donor cassette vectors used for the 4- cassette cloning strategy. C. The curved dotted lines indicate the overhangs that anneal during the ligation reaction. D. Overview of the assembly protocol. For a detailed protocol, see the Materials and Methods section. E. Summary of available promoter, gene tag and 3’UTR donor cassette plasmids.

    Article Snippet: Cloning To generate the expression vector pXF87, the two SapI restriction sites in pCFJ350 (Frøkjær-Jensen et al., 2012) were mutated using Q5 Site-Directed Mutagenesis (New England Biolabs) with the oligo pairs XF30F/XF30R and XF31F/XF31R.

    Techniques: Plasmid Preparation, Derivative Assay, Selection, Marker, Transgenic Assay, Clone Assay, Ligation

    SapTrap assembly of MosSCI targeting vectors using the four-cassette system. A. The MosSCI targeting vector pXF87 was derived from pCFJ350 by mutating two SapI restriction sites (indicated by arrowheads in the “Left” (L) and “Right” (R) homology arms) and introducing two SapI sites (blue text) between the XhoI and SpeI sites (green text). SapI cleavage sites are in red text. The SapI recognition sites are oriented such that upon digestion they are removed from the vector backbone. The cbr- unc-119 gene is used as a positive selection marker to facilitate the identification of transgenic animals. B. Design of the donor cassette vectors used for the 4-cassette cloning strategy. C. The curved dotted lines indicate the overhangs that anneal during the ligation reaction. D. Overview of the assembly protocol. For a detailed protocol, see the Materials and Methods section. E. Summary of available promoter, gene tag and 3′UTR donor cassette plasmids.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors

    doi: 10.1534/g3.119.400822

    Figure Lengend Snippet: SapTrap assembly of MosSCI targeting vectors using the four-cassette system. A. The MosSCI targeting vector pXF87 was derived from pCFJ350 by mutating two SapI restriction sites (indicated by arrowheads in the “Left” (L) and “Right” (R) homology arms) and introducing two SapI sites (blue text) between the XhoI and SpeI sites (green text). SapI cleavage sites are in red text. The SapI recognition sites are oriented such that upon digestion they are removed from the vector backbone. The cbr- unc-119 gene is used as a positive selection marker to facilitate the identification of transgenic animals. B. Design of the donor cassette vectors used for the 4-cassette cloning strategy. C. The curved dotted lines indicate the overhangs that anneal during the ligation reaction. D. Overview of the assembly protocol. For a detailed protocol, see the Materials and Methods section. E. Summary of available promoter, gene tag and 3′UTR donor cassette plasmids.

    Article Snippet: Cloning To generate the expression vector pXF87, the two SapI restriction sites in pCFJ350 ( ) were mutated using Q5 Site-Directed Mutagenesis (New England Biolabs (NEB)) with the oligo pairs XF30F/XF30R and XF31F/XF31R.

    Techniques: Plasmid Preparation, Derivative Assay, Selection, Marker, Transgenic Assay, Clone Assay, Ligation

    SapTrap assembly of MosSCI targeting vectors using the five-cassette system. A. Schematic of pXF87 and the donor cassettes following SapI digestion. The dotted lines indicate the overhangs that anneal during ligation. B. Summary of available promoter, gene tag and 3′UTR donor cassette plasmids for the five-cassette system.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors

    doi: 10.1534/g3.119.400822

    Figure Lengend Snippet: SapTrap assembly of MosSCI targeting vectors using the five-cassette system. A. Schematic of pXF87 and the donor cassettes following SapI digestion. The dotted lines indicate the overhangs that anneal during ligation. B. Summary of available promoter, gene tag and 3′UTR donor cassette plasmids for the five-cassette system.

    Article Snippet: Cloning To generate the expression vector pXF87, the two SapI restriction sites in pCFJ350 ( ) were mutated using Q5 Site-Directed Mutagenesis (New England Biolabs (NEB)) with the oligo pairs XF30F/XF30R and XF31F/XF31R.

    Techniques: Ligation

    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Journal: Current protocols in molecular biology

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo

    doi: 10.1002/cpmb.46

    Figure Lengend Snippet: Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Article Snippet: AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019).

    Techniques: Plasmid Preparation, Clone Assay, Construct

    Ube3b knockout in mice results in growth retardation, failure to thrive and an overall smaller brain. Related to the entire paper. (A) Gene targeting strategy of murine Ube3b . The domain structure of Ube3b, wild type, and targeted Ube3b alleles are depicted. Exons, loxP sites, and FLP-recombinase target sites are symbolized as black rectangles, red triangles, and green rectangles, respectively. Exon 7 of Ube3b is flanked by two loxP sites. Mice positive for the recombined allele were crossed with FLP deleter animals to remove the lacZ/neomycin selection cassette, and further crossed to Cre-driver lines, leading to removal of exon 7 conditionally or conventionally. (B, C, D) Validation of Ube3b gene targeting. +/+ wild type, rec/+ indicates heterozygous recombined mutant. (B) The result of long-range PCR using primers depicted as blue arrows in A. Band of the predicted size was observed only using genomic DNA purified from rec/+ mutant as a template. (C) The result of multiplex PCR using primers depicted as pink arrows in A. PCR using genomic DNA from targeted ES cells yields two bands (left lane), and PCR using wild type ES cells genomic DNA as a template results in one band. (D) Southern blotting analysis of genomic DNA purified from targeted ES cells. Genomic DNA isolated from control and ES cells carrying ‘Recombined’ allele of Ube3b was digested with SapI enzyme. The probe is indicated as a green line in A. The band at 8.0 kb represents the wild type allele, and the band at 6.6 kb represents the mutant allele. (E) Gross morphology of Ube3b +/+ (left) and Ube3b -/- (right) mice at 20 days post birth. (F) Brains of Ube3b +/+ (left) and Ube3b -/- (right) mice isolated 20 days after birth. (G) Results of Western blotting using P7 brain lysates from Ube3b +/+ (left), Ube3b +/- (middle), and Ube3b -/- (right) animals with an antibody raised against HECT domain of Ube3b. MEM Code staining (lower panel) shows comparable amounts of total protein in all lanes.

    Journal: bioRxiv

    Article Title: The Kaufman oculocerebrofacial syndrome protein Ube3b regulates synapse number by ubiquitinating Ppp3cc

    doi: 10.1101/672923

    Figure Lengend Snippet: Ube3b knockout in mice results in growth retardation, failure to thrive and an overall smaller brain. Related to the entire paper. (A) Gene targeting strategy of murine Ube3b . The domain structure of Ube3b, wild type, and targeted Ube3b alleles are depicted. Exons, loxP sites, and FLP-recombinase target sites are symbolized as black rectangles, red triangles, and green rectangles, respectively. Exon 7 of Ube3b is flanked by two loxP sites. Mice positive for the recombined allele were crossed with FLP deleter animals to remove the lacZ/neomycin selection cassette, and further crossed to Cre-driver lines, leading to removal of exon 7 conditionally or conventionally. (B, C, D) Validation of Ube3b gene targeting. +/+ wild type, rec/+ indicates heterozygous recombined mutant. (B) The result of long-range PCR using primers depicted as blue arrows in A. Band of the predicted size was observed only using genomic DNA purified from rec/+ mutant as a template. (C) The result of multiplex PCR using primers depicted as pink arrows in A. PCR using genomic DNA from targeted ES cells yields two bands (left lane), and PCR using wild type ES cells genomic DNA as a template results in one band. (D) Southern blotting analysis of genomic DNA purified from targeted ES cells. Genomic DNA isolated from control and ES cells carrying ‘Recombined’ allele of Ube3b was digested with SapI enzyme. The probe is indicated as a green line in A. The band at 8.0 kb represents the wild type allele, and the band at 6.6 kb represents the mutant allele. (E) Gross morphology of Ube3b +/+ (left) and Ube3b -/- (right) mice at 20 days post birth. (F) Brains of Ube3b +/+ (left) and Ube3b -/- (right) mice isolated 20 days after birth. (G) Results of Western blotting using P7 brain lysates from Ube3b +/+ (left), Ube3b +/- (middle), and Ube3b -/- (right) animals with an antibody raised against HECT domain of Ube3b. MEM Code staining (lower panel) shows comparable amounts of total protein in all lanes.

    Article Snippet: Southern blotting Restriction digest of 10 μg of genomic DNA isolated from ES cells was held using 10U of SapI restriction enzyme (NEB) in 37°C for 16 hours.

    Techniques: Knock-Out, Mouse Assay, Selection, Mutagenesis, Polymerase Chain Reaction, Purification, Multiplex Assay, Southern Blot, Isolation, Western Blot, Staining