kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc kb dna ladder
    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb <t>DNA</t> Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used
    Kb Dna Ladder, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    Images

    1) Product Images from "Design, Expression and Purification of Strongyloides stercoralis IgG4 Immunoreactive Protein (NIE) in Escherichia coli"

    Article Title: Design, Expression and Purification of Strongyloides stercoralis IgG4 Immunoreactive Protein (NIE) in Escherichia coli

    Journal: Iranian Journal of Parasitology

    doi: 10.18502/ijpa.v15i3.4198

    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used
    Figure Legend Snippet: Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used

    Techniques Used: Negative Control, Positive Control, Sequencing

    kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc kb dna ladder
    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb <t>DNA</t> Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used
    Kb Dna Ladder, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kb dna ladder - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Design, Expression and Purification of Strongyloides stercoralis IgG4 Immunoreactive Protein (NIE) in Escherichia coli"

    Article Title: Design, Expression and Purification of Strongyloides stercoralis IgG4 Immunoreactive Protein (NIE) in Escherichia coli

    Journal: Iranian Journal of Parasitology

    doi: 10.18502/ijpa.v15i3.4198

    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used
    Figure Legend Snippet: Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used

    Techniques Used: Negative Control, Positive Control, Sequencing

    kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc kb dna ladder
    Colony PCR of the colonies; Lanes 1–8: eight selected colonies from the plate; Lane 9: 1 kb <t>DNA</t> <t>Ladder</t>
    Kb Dna Ladder, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    1) Product Images from "Investigating the Efficiency of Recombinant FliC-Loaded Bacillus subtilis Spores in Mice Immunization against Salmonella enterica Serovar Typhi"

    Article Title: Investigating the Efficiency of Recombinant FliC-Loaded Bacillus subtilis Spores in Mice Immunization against Salmonella enterica Serovar Typhi

    Journal: Iranian Journal of Public Health

    doi: 10.18502/ijph.v50i7.6638

    Colony PCR of the colonies; Lanes 1–8: eight selected colonies from the plate; Lane 9: 1 kb DNA Ladder
    Figure Legend Snippet: Colony PCR of the colonies; Lanes 1–8: eight selected colonies from the plate; Lane 9: 1 kb DNA Ladder

    Techniques Used:

    kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc kb dna ladder
    Kb Dna Ladder, supplied by Gold Biotechnology Inc, 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/kb dna ladder/product/Gold Biotechnology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    1 kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc 1 kb dna ladder
    Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp <t>DNA</t> ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies
    1 Kb Dna Ladder, supplied by Gold Biotechnology Inc, 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/1 kb dna ladder/product/Gold Biotechnology Inc
    Average 94 stars, based on 1 article reviews
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    1 kb dna ladder - by Bioz Stars, 2023-02
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    Images

    1) Product Images from "Pyrite cloning: a single tube and programmed reaction cloning with restriction enzymes"

    Article Title: Pyrite cloning: a single tube and programmed reaction cloning with restriction enzymes

    Journal: Plant Methods

    doi: 10.1186/s13007-018-0359-7

    Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp DNA ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies
    Figure Legend Snippet: Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp DNA ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies

    Techniques Used: Clone Assay, Marker, Colony Assay, Transformation Assay, Plasmid Preparation, Ligation

    Multiple applications of Pyrite cloning. a Circularization/ligation of a linear DNA fragment. A PCR amplified linear vector can be cut and ligated using the Pyrite reaction. pLacZi was amplified with primers MF143 and MF144 and put in the Pyrite reaction with HindIII-HF, restoring the HindIII site in pLacZi. MF41 and MF126 are PCR primers used to test successful cloning. Three of the eight colonies tested were positive and marked by white stars. b Insertion of a DNA fragment into multiple destination vectors in a single reaction. The transformed cells can be plated on different antibiotic containing media to select for the desired vector. The gel image on the left shows the insertion of eGFP into two vectors simultaneously; the image on the right shows the insertion of gene31413 CDS into two vectors simultaneously. Red star indicates negative control (no template). Green stars indicate the vector pMerlin, which confers AmpR. Orange stars indicate the vector pSanFran, which confers SmR. “m” indicates marker lane (Goldbio 100 bp DNA ladder). c Restriction digestion of extracted plasmid DNA by EcoRI and SalI to release the cloned fragment. Proper insert and vector size confirmed colony PCR results. Arrows indicates the released insert of 817 bp and 727 bp respectively. Green star indicates vector that conferred AmpR (pMerlin). Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances. Two successful examples are shown with 67–100% efficiency (positive PCR products marked by white stars). e A vector with an insert can revert to the empty vector by releasing the insert with the Pyrite reaction. Gene31413 CDS was removed from the pCR™8/ GW / TOPO ® vector with EcoRI-HF. All colonies screened contain the empty vector without an insert (blue stars)
    Figure Legend Snippet: Multiple applications of Pyrite cloning. a Circularization/ligation of a linear DNA fragment. A PCR amplified linear vector can be cut and ligated using the Pyrite reaction. pLacZi was amplified with primers MF143 and MF144 and put in the Pyrite reaction with HindIII-HF, restoring the HindIII site in pLacZi. MF41 and MF126 are PCR primers used to test successful cloning. Three of the eight colonies tested were positive and marked by white stars. b Insertion of a DNA fragment into multiple destination vectors in a single reaction. The transformed cells can be plated on different antibiotic containing media to select for the desired vector. The gel image on the left shows the insertion of eGFP into two vectors simultaneously; the image on the right shows the insertion of gene31413 CDS into two vectors simultaneously. Red star indicates negative control (no template). Green stars indicate the vector pMerlin, which confers AmpR. Orange stars indicate the vector pSanFran, which confers SmR. “m” indicates marker lane (Goldbio 100 bp DNA ladder). c Restriction digestion of extracted plasmid DNA by EcoRI and SalI to release the cloned fragment. Proper insert and vector size confirmed colony PCR results. Arrows indicates the released insert of 817 bp and 727 bp respectively. Green star indicates vector that conferred AmpR (pMerlin). Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances. Two successful examples are shown with 67–100% efficiency (positive PCR products marked by white stars). e A vector with an insert can revert to the empty vector by releasing the insert with the Pyrite reaction. Gene31413 CDS was removed from the pCR™8/ GW / TOPO ® vector with EcoRI-HF. All colonies screened contain the empty vector without an insert (blue stars)

    Techniques Used: Clone Assay, Ligation, Amplification, Plasmid Preparation, Transformation Assay, Negative Control, Marker

    1 kb dna ladder  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc 1 kb dna ladder
    PL Vagabond primers discriminate between P. larvae and other closely related bacteria. (A and B) Paenibacillus-specific primers were used to attempt PCR amplification of a region of the genome found in various control strains as well as field isolates. TAQ (New England BioLabs) PCR was performed according to the manufacturer's instructions with an annealing temperature of 64°C (A) or 60°C (B). Lanes were loaded identically for the two gels as follows (the bacterial template is indicated): 1, B. laterosporus 40A1 (ATCC 9141); 2, Br1; 3, Br2; 4, Br3; 5, Br4; 6, Br5; 7, Br6; 8, Br7; 9, Br9; 10, Br14; 11, B. brevis (ATCC 8246); 12, P. larvae (ATCC 9545); 13, PL309; 14, PL325; 15, PL335; 16, PL337; 17, PL338; 18, PL344; 19, PL345; 20, PL357; 21, Paenibacillus sp. (BGSC 35A1); 22, P. alvei (BGSC 33A1); 23, P. polymyxa (BGSC 25A2); 24, negative control. The black line indicates a separate gel. The ladder included at the far left lane of each gel is a <t>1-kb</t> <t>DNA</t> <t>ladder</t> (Gold Biotechnology), with the top bright band representing 3 kb and the bottom bright band representing 1 kb.
    1 Kb Dna Ladder, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "A PCR-Based Method for Distinguishing between Two Common Beehive Bacteria, Paenibacillus larvae and Brevibacillus laterosporus"

    Article Title: A PCR-Based Method for Distinguishing between Two Common Beehive Bacteria, Paenibacillus larvae and Brevibacillus laterosporus

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.01886-18

    PL Vagabond primers discriminate between P. larvae and other closely related bacteria. (A and B) Paenibacillus-specific primers were used to attempt PCR amplification of a region of the genome found in various control strains as well as field isolates. TAQ (New England BioLabs) PCR was performed according to the manufacturer's instructions with an annealing temperature of 64°C (A) or 60°C (B). Lanes were loaded identically for the two gels as follows (the bacterial template is indicated): 1, B. laterosporus 40A1 (ATCC 9141); 2, Br1; 3, Br2; 4, Br3; 5, Br4; 6, Br5; 7, Br6; 8, Br7; 9, Br9; 10, Br14; 11, B. brevis (ATCC 8246); 12, P. larvae (ATCC 9545); 13, PL309; 14, PL325; 15, PL335; 16, PL337; 17, PL338; 18, PL344; 19, PL345; 20, PL357; 21, Paenibacillus sp. (BGSC 35A1); 22, P. alvei (BGSC 33A1); 23, P. polymyxa (BGSC 25A2); 24, negative control. The black line indicates a separate gel. The ladder included at the far left lane of each gel is a 1-kb DNA ladder (Gold Biotechnology), with the top bright band representing 3 kb and the bottom bright band representing 1 kb.
    Figure Legend Snippet: PL Vagabond primers discriminate between P. larvae and other closely related bacteria. (A and B) Paenibacillus-specific primers were used to attempt PCR amplification of a region of the genome found in various control strains as well as field isolates. TAQ (New England BioLabs) PCR was performed according to the manufacturer's instructions with an annealing temperature of 64°C (A) or 60°C (B). Lanes were loaded identically for the two gels as follows (the bacterial template is indicated): 1, B. laterosporus 40A1 (ATCC 9141); 2, Br1; 3, Br2; 4, Br3; 5, Br4; 6, Br5; 7, Br6; 8, Br7; 9, Br9; 10, Br14; 11, B. brevis (ATCC 8246); 12, P. larvae (ATCC 9545); 13, PL309; 14, PL325; 15, PL335; 16, PL337; 17, PL338; 18, PL344; 19, PL345; 20, PL357; 21, Paenibacillus sp. (BGSC 35A1); 22, P. alvei (BGSC 33A1); 23, P. polymyxa (BGSC 25A2); 24, negative control. The black line indicates a separate gel. The ladder included at the far left lane of each gel is a 1-kb DNA ladder (Gold Biotechnology), with the top bright band representing 3 kb and the bottom bright band representing 1 kb.

    Techniques Used: Amplification, Negative Control

    Brevibacillus-specific primers for the rpoB gene discriminate between Paenibacillus and Brevibacillus bacteria and verify that field isolates (BL) are Brevibacillus laterosporus. Brevibacillus-specific primers (BL rpoB) were used to attempt PCR amplification of the rpoB gene found in B. laterosporus from various control strains as well as field isolates (BL). TAQ (New England BioLabs) PCR was performed according to the manufacturer's instructions with an annealing temperature of 69.1°C. Lanes were loaded as follows (the bacterial template is indicated): 1, B. laterosporus 40A1 (ATCC 9141); 2, B. brevis (ATCC 8246); 3, P. larvae (ATCC 9545); 4, P. alvei (BGSC 33A1); 5, P. polymyxa (BGSC 25A2); 6, Paenibacillus sp. (BGSC 35A1); 7, Br1; 8, Br3; 9, Br4; 10, Br5; 11, Br6; 12, Br12; 13, Brevibacillus sp.; 14, PL357. The black line indicates a separate gel. The ladder included in the far-left lane of each gel is a 1-kb Plus DNA ladder (Gold Biotechnology), with the top bright band representing 3 kb and the bottom bright band representing 1 kb.
    Figure Legend Snippet: Brevibacillus-specific primers for the rpoB gene discriminate between Paenibacillus and Brevibacillus bacteria and verify that field isolates (BL) are Brevibacillus laterosporus. Brevibacillus-specific primers (BL rpoB) were used to attempt PCR amplification of the rpoB gene found in B. laterosporus from various control strains as well as field isolates (BL). TAQ (New England BioLabs) PCR was performed according to the manufacturer's instructions with an annealing temperature of 69.1°C. Lanes were loaded as follows (the bacterial template is indicated): 1, B. laterosporus 40A1 (ATCC 9141); 2, B. brevis (ATCC 8246); 3, P. larvae (ATCC 9545); 4, P. alvei (BGSC 33A1); 5, P. polymyxa (BGSC 25A2); 6, Paenibacillus sp. (BGSC 35A1); 7, Br1; 8, Br3; 9, Br4; 10, Br5; 11, Br6; 12, Br12; 13, Brevibacillus sp.; 14, PL357. The black line indicates a separate gel. The ladder included in the far-left lane of each gel is a 1-kb Plus DNA ladder (Gold Biotechnology), with the top bright band representing 3 kb and the bottom bright band representing 1 kb.

    Techniques Used: Amplification

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    Gold Biotechnology Inc kb dna ladder
    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb <t>DNA</t> Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used
    Kb Dna Ladder, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gold Biotechnology Inc 1 kb dna ladder
    Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp <t>DNA</t> ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies
    1 Kb Dna Ladder, supplied by Gold Biotechnology Inc, 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/1 kb dna ladder/product/Gold Biotechnology Inc
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    Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used

    Journal: Iranian Journal of Parasitology

    Article Title: Design, Expression and Purification of Strongyloides stercoralis IgG4 Immunoreactive Protein (NIE) in Escherichia coli

    doi: 10.18502/ijpa.v15i3.4198

    Figure Lengend Snippet: Colony PCR of the grown colonies on LB agar supplemented with Kanamycin. Lanes 1–4: Four different colonies grown on the LB agar; Lane 5: Negative control. Lane 6: Positive control; M: 1 kb DNA Ladder. In negative control, all materials of the PCR reaction were as the same as the test samples but the template DNA, which was the genomic DNA of E. coli BL21 (DE3) strain. In positive control, another DNA sequence (with a size of about 2 kb) with its primers was used

    Article Snippet: PCR master mix and 1 kb DNA Ladder were purchased from GoldBio(China).

    Techniques: Negative Control, Positive Control, Sequencing

    Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp DNA ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies

    Journal: Plant Methods

    Article Title: Pyrite cloning: a single tube and programmed reaction cloning with restriction enzymes

    doi: 10.1186/s13007-018-0359-7

    Figure Lengend Snippet: Determination of Pyrite cloning efficiency using colony PCR and blue-white screen. a Examples of colony PCR from three different reactions (i, ii, and iii), showing about 55% colonies (10/18; star) with the expected insert. “m” indicates marker lane (Goldbio 100 bp DNA ladder). Reaction i screened for insertion of F. vesca gene30478 CDS into pB42AD, reaction ii screened for insertion of F. vesca gene30478 CDS into pLexA, and reaction iii screened for insertion of F. vesca gene25060 CDS into pB42AD. b – g Blue-white colony screening used to determine the efficiency of the multiple applications of Pyrite cloning. Colonies are formed on antibiotic containing LB agar plates spread with X-gal and IPTG. b Control experiment showing 100% blue colonies after transformation of vector pUC19. c pUC19 was put into the Pyrite reaction to determine background re-ligation in the absence of DNA fragments. A smaller number of blue colonies was observed. d Pyrite reaction to insert F. vesca gene25060 CDS into pUC19. 100% white colonies were observed. e Pyrite cloning of gene25060 into pUC19 with SalI and BamHI-HF, a heat-resistant restriction enzyme, yielded 83% white colonies. f Simultaneous cloning of a F. vesca gene31413 into two vectors, pUC19 and pSanFran, in the same tube. The result of cloning gene31413 into pUC19 is shown. g Swapping eGFP from pSanFran to pUC19 with the Pyrite reaction, yielded 49.3% white colonies

    Article Snippet: Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances.

    Techniques: Clone Assay, Marker, Colony Assay, Transformation Assay, Plasmid Preparation, Ligation

    Multiple applications of Pyrite cloning. a Circularization/ligation of a linear DNA fragment. A PCR amplified linear vector can be cut and ligated using the Pyrite reaction. pLacZi was amplified with primers MF143 and MF144 and put in the Pyrite reaction with HindIII-HF, restoring the HindIII site in pLacZi. MF41 and MF126 are PCR primers used to test successful cloning. Three of the eight colonies tested were positive and marked by white stars. b Insertion of a DNA fragment into multiple destination vectors in a single reaction. The transformed cells can be plated on different antibiotic containing media to select for the desired vector. The gel image on the left shows the insertion of eGFP into two vectors simultaneously; the image on the right shows the insertion of gene31413 CDS into two vectors simultaneously. Red star indicates negative control (no template). Green stars indicate the vector pMerlin, which confers AmpR. Orange stars indicate the vector pSanFran, which confers SmR. “m” indicates marker lane (Goldbio 100 bp DNA ladder). c Restriction digestion of extracted plasmid DNA by EcoRI and SalI to release the cloned fragment. Proper insert and vector size confirmed colony PCR results. Arrows indicates the released insert of 817 bp and 727 bp respectively. Green star indicates vector that conferred AmpR (pMerlin). Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances. Two successful examples are shown with 67–100% efficiency (positive PCR products marked by white stars). e A vector with an insert can revert to the empty vector by releasing the insert with the Pyrite reaction. Gene31413 CDS was removed from the pCR™8/ GW / TOPO ® vector with EcoRI-HF. All colonies screened contain the empty vector without an insert (blue stars)

    Journal: Plant Methods

    Article Title: Pyrite cloning: a single tube and programmed reaction cloning with restriction enzymes

    doi: 10.1186/s13007-018-0359-7

    Figure Lengend Snippet: Multiple applications of Pyrite cloning. a Circularization/ligation of a linear DNA fragment. A PCR amplified linear vector can be cut and ligated using the Pyrite reaction. pLacZi was amplified with primers MF143 and MF144 and put in the Pyrite reaction with HindIII-HF, restoring the HindIII site in pLacZi. MF41 and MF126 are PCR primers used to test successful cloning. Three of the eight colonies tested were positive and marked by white stars. b Insertion of a DNA fragment into multiple destination vectors in a single reaction. The transformed cells can be plated on different antibiotic containing media to select for the desired vector. The gel image on the left shows the insertion of eGFP into two vectors simultaneously; the image on the right shows the insertion of gene31413 CDS into two vectors simultaneously. Red star indicates negative control (no template). Green stars indicate the vector pMerlin, which confers AmpR. Orange stars indicate the vector pSanFran, which confers SmR. “m” indicates marker lane (Goldbio 100 bp DNA ladder). c Restriction digestion of extracted plasmid DNA by EcoRI and SalI to release the cloned fragment. Proper insert and vector size confirmed colony PCR results. Arrows indicates the released insert of 817 bp and 727 bp respectively. Green star indicates vector that conferred AmpR (pMerlin). Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances. Two successful examples are shown with 67–100% efficiency (positive PCR products marked by white stars). e A vector with an insert can revert to the empty vector by releasing the insert with the Pyrite reaction. Gene31413 CDS was removed from the pCR™8/ GW / TOPO ® vector with EcoRI-HF. All colonies screened contain the empty vector without an insert (blue stars)

    Article Snippet: Orange star indicates vector that conferred SpecR (pSanfran). m indicates Goldbio 1 kb DNA ladder. d Insert swapping between vectors with different antibiotic resistances.

    Techniques: Clone Assay, Ligation, Amplification, Plasmid Preparation, Transformation Assay, Negative Control, Marker