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  • 99
    Thermo Fisher electromax dh10b cells
    Generation of a BAC of E1-deleted Ad3 encoding the eGFP reporter gene. (A) The plasmid pBSAd3LRzeo contains a zeocin selection cassette flanked by a 468-bp fragment of the Ad3 left end sequence and a 528-bp fragment of the Ad3 right end sequence. (B) Transfer of the Not IB– Hin dIII fragment containing the Ad3LRzeo cassette to the single copy pKSB2 plasmid. (C) Transformation of <t>DH10B</t> bacteria expressing the phage lambda recombinases allows homologous recombination between the Sal I-linearized pKSB2Ad3LRzeo and Ad3 genomic DNA, resulting in pKSBAd3wt. (D) Subsequent homologous recombination between pKSBAd3wt and a CMV-eGFP/zeo cassette flanked by short homologous sequences of 40 bp results in pKSB2 Ad3CMV-eGFP, which contains the eGFP expression cassette in reverse orientation. (E) Release of the viral genome by the flanking unique Mlu I endonuclease sites followed by transfection of helper 911-Ad3E1B cells yielding Ad3CMV-eGFP. (For details of the procedure, see Materials and methods .)
    Electromax Dh10b Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher e coli dh10b
    mTn5 transposon mutagenesis using an IPTG-controlled conditional suicide plasmid. a Diagram of plasmid pSNC-mTn5. pSNC-mTn5 is a derivative of pMMB-repF that contains a Kan R -tagged mTn5, a lac promoter-controlled hyperactive transposase gene ( tnp H ), and a sacB counter selection marker (with its own promoter). OE and IE are outside and inside ends of the mTn5. b Plasmid and transposon retention frequencies in E. coli <t>DH10B.</t> A “+” symbol for IPTG indicates that the inducer was added to the liquid culture, and a “+” symbol for Suc, Cam, and Kan indicates that the chemicals were added to the plates. Black columns represent plasmid retention frequencies, and the blue column represents Tn5 retention frequency. Results were average of three independent experiments, and bars represent mean ± SD (*p
    E Coli Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1302 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher e coli strain dh10b
    Strategy for constructing a self-excisable RhCMV BAC. (A) Cloning of the full-length RhCMV genome into a BAC vector by Cre/lox recombination. The RhCMV genome structure with expansion of the US1-to-US3 region of recombinant viruses is diagrammed. The open box represents the internal junction between L and S components of the RhCMV genome. Cre/lox recombination was performed in Telo-RF cells. Recombinant clones containing the BAC vector in the viral genome (vRhCMV/BAC-EGFP) were plaque purified, and circular-form viral DNA was transformed into E. coli strain <t>DH10B</t> for plasmid isolation. (B) Substitution of the cre ORF for the egfp ORF by allelic exchange in E. coli . Only a portion of each plasmid is illustrated. The homologous flanking regions and the cre ORF of the recombination vector, pWC212, are illustrated. UL, unique sequences of the L component; US, unique sequences of the S component; PSV40, SV40 promoter; poly A, SV40 polyadenylation signal.
    E Coli Strain Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 378 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher e coli dh10b cells
    Stability of the BAC pBACBHV37 over multiple generations in E. coli <t>DH10B.</t> DH10B cells containing pBACBHV37 were serially grown for 5, 10, and 20 days. BAC DNA was prepared from these cultures, and the restriction enzyme profiles were compared by FIGE with 1% agarose. DNA fragments were stained with ethidium bromide and photographed. Standard molecular sizes (in base pairs) are given.
    E Coli Dh10b Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 498 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher megax dh10b t1r electrocomp cells
    Stability of the BAC pBACBHV37 over multiple generations in E. coli <t>DH10B.</t> DH10B cells containing pBACBHV37 were serially grown for 5, 10, and 20 days. BAC DNA was prepared from these cultures, and the restriction enzyme profiles were compared by FIGE with 1% agarose. DNA fragments were stained with ethidium bromide and photographed. Standard molecular sizes (in base pairs) are given.
    Megax Dh10b T1r Electrocomp Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher max efficiency dh10b competent cells
    Drop assay of the four clones resistant to UV radiation to test perchlorate resistance. <t>DH10B</t> pBlueScript SK (+) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid medium supplemented with perchlorate 125 mM. An additional test was performed combining UVB (312 nm) radiation exposure during 80 s at 2.26 W m –2 and perchlorate (125 mM). A negative control without irradiation (–) was performed to verify the normal growth of the clones. Each experiment was performed at least three times using independent cultures.
    Max Efficiency Dh10b Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher dh10b competent cells
    Drop assay of the four clones resistant to UV radiation to test perchlorate resistance. <t>DH10B</t> pBlueScript SK (+) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid medium supplemented with perchlorate 125 mM. An additional test was performed combining UVB (312 nm) radiation exposure during 80 s at 2.26 W m –2 and perchlorate (125 mM). A negative control without irradiation (–) was performed to verify the normal growth of the clones. Each experiment was performed at least three times using independent cultures.
    Dh10b Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 263 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher electrocompetent dh10b cells
    A typical chimeric-GFP solubility screening experiment. ( A ) white light image, ( B ) transilluminating UV (365 nm) image of an agar plate used to select GFP-positive colonies in the solubility screen. <t>Electrocompetent</t> <t>DH10B</t> cells were transformed with a
    Electrocompetent Dh10b Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher electrocompetent escherichia coli dh10b
    A typical chimeric-GFP solubility screening experiment. ( A ) white light image, ( B ) transilluminating UV (365 nm) image of an agar plate used to select GFP-positive colonies in the solubility screen. <t>Electrocompetent</t> <t>DH10B</t> cells were transformed with a
    Electrocompetent Escherichia Coli Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad e coli dh10b
    Metalloid resistance conferred to Escherichia coli <t>DH10B</t> by metagenomic clone pA7 and pA12. Resistance to a Sodium arsenite, b Sodium arsenate and c Potassium antimony tartrate was examined with E. coli DH10B + pHB201(control) ( filled circle ), E. coli DH10B + pA7 ( filled triangle ) and E. coli DH10B + pA12 ( filled square ). Experiments performed in triplicates, values are averages and standard deviations are shown as error bars
    E Coli Dh10b, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad escherichia coli dh10b cells
    Metalloid resistance conferred to Escherichia coli <t>DH10B</t> by metagenomic clone pA7 and pA12. Resistance to a Sodium arsenite, b Sodium arsenate and c Potassium antimony tartrate was examined with E. coli DH10B + pHB201(control) ( filled circle ), E. coli DH10B + pA7 ( filled triangle ) and E. coli DH10B + pA12 ( filled square ). Experiments performed in triplicates, values are averages and standard deviations are shown as error bars
    Escherichia Coli Dh10b Cells, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher electrocompetent escherichia coli strain dh10b
    Metalloid resistance conferred to Escherichia coli <t>DH10B</t> by metagenomic clone pA7 and pA12. Resistance to a Sodium arsenite, b Sodium arsenate and c Potassium antimony tartrate was examined with E. coli DH10B + pHB201(control) ( filled circle ), E. coli DH10B + pA7 ( filled triangle ) and E. coli DH10B + pA12 ( filled square ). Experiments performed in triplicates, values are averages and standard deviations are shown as error bars
    Electrocompetent Escherichia Coli Strain Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Generation of a BAC of E1-deleted Ad3 encoding the eGFP reporter gene. (A) The plasmid pBSAd3LRzeo contains a zeocin selection cassette flanked by a 468-bp fragment of the Ad3 left end sequence and a 528-bp fragment of the Ad3 right end sequence. (B) Transfer of the Not IB– Hin dIII fragment containing the Ad3LRzeo cassette to the single copy pKSB2 plasmid. (C) Transformation of DH10B bacteria expressing the phage lambda recombinases allows homologous recombination between the Sal I-linearized pKSB2Ad3LRzeo and Ad3 genomic DNA, resulting in pKSBAd3wt. (D) Subsequent homologous recombination between pKSBAd3wt and a CMV-eGFP/zeo cassette flanked by short homologous sequences of 40 bp results in pKSB2 Ad3CMV-eGFP, which contains the eGFP expression cassette in reverse orientation. (E) Release of the viral genome by the flanking unique Mlu I endonuclease sites followed by transfection of helper 911-Ad3E1B cells yielding Ad3CMV-eGFP. (For details of the procedure, see Materials and methods .)

    Journal: Virology

    Article Title: The nucleotide sequence and a first generation gene transfer vector of species B human adenovirus serotype 3

    doi: 10.1016/j.virol.2005.08.024

    Figure Lengend Snippet: Generation of a BAC of E1-deleted Ad3 encoding the eGFP reporter gene. (A) The plasmid pBSAd3LRzeo contains a zeocin selection cassette flanked by a 468-bp fragment of the Ad3 left end sequence and a 528-bp fragment of the Ad3 right end sequence. (B) Transfer of the Not IB– Hin dIII fragment containing the Ad3LRzeo cassette to the single copy pKSB2 plasmid. (C) Transformation of DH10B bacteria expressing the phage lambda recombinases allows homologous recombination between the Sal I-linearized pKSB2Ad3LRzeo and Ad3 genomic DNA, resulting in pKSBAd3wt. (D) Subsequent homologous recombination between pKSBAd3wt and a CMV-eGFP/zeo cassette flanked by short homologous sequences of 40 bp results in pKSB2 Ad3CMV-eGFP, which contains the eGFP expression cassette in reverse orientation. (E) Release of the viral genome by the flanking unique Mlu I endonuclease sites followed by transfection of helper 911-Ad3E1B cells yielding Ad3CMV-eGFP. (For details of the procedure, see Materials and methods .)

    Article Snippet: In order to generate pKSB2Ad3wt, which contained the complete Ad3 genome, homologous recombination in DH10B (Invitrogen) was performed.

    Techniques: BAC Assay, Plasmid Preparation, Selection, Sequencing, Transformation Assay, Expressing, Homologous Recombination, Transfection

    The construction method of Os_GH-pmKate2-N mammalian expression vector. The Os_GH-pTZ57R/T cloning vector and pmKate2-N expression vector were digested with SacI and SacII restriction enzymes to make sticky ends in both Os_GH cDNA fragment and pmKate2-N vector. The digested Os_GH cDNA fragment was ligated into the digested pmKate2-N vector. Finally, the constructed Os_GH-pmKate2-N vector was transformed into DH10B cells for amplification (adapted by authors).

    Journal: Journal of Genetic Engineering & Biotechnology

    Article Title: Construction of ovine GH-pmKate2N expression vector and its uptake by ovine spermatozoa using different methods

    doi: 10.1016/j.jgeb.2017.04.001

    Figure Lengend Snippet: The construction method of Os_GH-pmKate2-N mammalian expression vector. The Os_GH-pTZ57R/T cloning vector and pmKate2-N expression vector were digested with SacI and SacII restriction enzymes to make sticky ends in both Os_GH cDNA fragment and pmKate2-N vector. The digested Os_GH cDNA fragment was ligated into the digested pmKate2-N vector. Finally, the constructed Os_GH-pmKate2-N vector was transformed into DH10B cells for amplification (adapted by authors).

    Article Snippet: The constructed Os_GH-pmKate2-N vector was transformed into DH10B cells according to the kit instructions (Fermentas, USA) and extracted using GeneJET plasmid Miniprep kit.

    Techniques: Expressing, Plasmid Preparation, Clone Assay, Construct, Transformation Assay, Amplification

    mTn5 transposon mutagenesis using an IPTG-controlled conditional suicide plasmid. a Diagram of plasmid pSNC-mTn5. pSNC-mTn5 is a derivative of pMMB-repF that contains a Kan R -tagged mTn5, a lac promoter-controlled hyperactive transposase gene ( tnp H ), and a sacB counter selection marker (with its own promoter). OE and IE are outside and inside ends of the mTn5. b Plasmid and transposon retention frequencies in E. coli DH10B. A “+” symbol for IPTG indicates that the inducer was added to the liquid culture, and a “+” symbol for Suc, Cam, and Kan indicates that the chemicals were added to the plates. Black columns represent plasmid retention frequencies, and the blue column represents Tn5 retention frequency. Results were average of three independent experiments, and bars represent mean ± SD (*p

    Journal: BMC Microbiology

    Article Title: Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid

    doi: 10.1186/s12866-018-1319-0

    Figure Lengend Snippet: mTn5 transposon mutagenesis using an IPTG-controlled conditional suicide plasmid. a Diagram of plasmid pSNC-mTn5. pSNC-mTn5 is a derivative of pMMB-repF that contains a Kan R -tagged mTn5, a lac promoter-controlled hyperactive transposase gene ( tnp H ), and a sacB counter selection marker (with its own promoter). OE and IE are outside and inside ends of the mTn5. b Plasmid and transposon retention frequencies in E. coli DH10B. A “+” symbol for IPTG indicates that the inducer was added to the liquid culture, and a “+” symbol for Suc, Cam, and Kan indicates that the chemicals were added to the plates. Black columns represent plasmid retention frequencies, and the blue column represents Tn5 retention frequency. Results were average of three independent experiments, and bars represent mean ± SD (*p

    Article Snippet: E. coli DH10B was purchased from Invitrogen.

    Techniques: Mutagenesis, Plasmid Preparation, Selection, Marker, Chick Chorioallantoic Membrane Assay

    Generation of a P. aeruginosa insertion library with pSNC-mTn5ME. a Diagram of pSNC-mTn5ME, a derivative of pSNC-mTn5 that has MEs instead of OE and IE at the termini of mTn5. b Plasmid and transposon retention frequencies in E. coli DH10B. Results were average of three independent experiments, and bars represent mean ± SD (* p

    Journal: BMC Microbiology

    Article Title: Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid

    doi: 10.1186/s12866-018-1319-0

    Figure Lengend Snippet: Generation of a P. aeruginosa insertion library with pSNC-mTn5ME. a Diagram of pSNC-mTn5ME, a derivative of pSNC-mTn5 that has MEs instead of OE and IE at the termini of mTn5. b Plasmid and transposon retention frequencies in E. coli DH10B. Results were average of three independent experiments, and bars represent mean ± SD (* p

    Article Snippet: E. coli DH10B was purchased from Invitrogen.

    Techniques: Plasmid Preparation

    IPTG-controlled conditional suicide plasmids. a Plasmid pMMB208 and its conditional-suicide derivatives. pMMB208 contains an RSF1010 oriV for replication and an oriT for conjugation. Genes repA , mobA / repB and repC encode proteins required for plasmid replication, and repF encodes a transcription repressor that binds promoter P4. pMMB208 also has Cam R and lacI Q genes and a P tac promoter. Plasmid pMMB- repF is a derivative of pMMB208 that has a second copy of the repF gene inserted downstream of P tac . Plasmids pMMB- repA K42A and pMMB- repA D139A have a dominant-negative repA mutant gene, either K42A or D139A, inserted downstream of P tac . b Amount of E. coli DH10B cells retaining the indicated plasmids after 24 h growth in the absence of antibiotics, either with or without IPTG induction. Results were average of three independent experiments, and bars represent mean ± SD (standard deviation). * p

    Journal: BMC Microbiology

    Article Title: Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid

    doi: 10.1186/s12866-018-1319-0

    Figure Lengend Snippet: IPTG-controlled conditional suicide plasmids. a Plasmid pMMB208 and its conditional-suicide derivatives. pMMB208 contains an RSF1010 oriV for replication and an oriT for conjugation. Genes repA , mobA / repB and repC encode proteins required for plasmid replication, and repF encodes a transcription repressor that binds promoter P4. pMMB208 also has Cam R and lacI Q genes and a P tac promoter. Plasmid pMMB- repF is a derivative of pMMB208 that has a second copy of the repF gene inserted downstream of P tac . Plasmids pMMB- repA K42A and pMMB- repA D139A have a dominant-negative repA mutant gene, either K42A or D139A, inserted downstream of P tac . b Amount of E. coli DH10B cells retaining the indicated plasmids after 24 h growth in the absence of antibiotics, either with or without IPTG induction. Results were average of three independent experiments, and bars represent mean ± SD (standard deviation). * p

    Article Snippet: E. coli DH10B was purchased from Invitrogen.

    Techniques: Plasmid Preparation, Conjugation Assay, Chick Chorioallantoic Membrane Assay, Dominant Negative Mutation, Mutagenesis, Standard Deviation

    Strategy for constructing a self-excisable RhCMV BAC. (A) Cloning of the full-length RhCMV genome into a BAC vector by Cre/lox recombination. The RhCMV genome structure with expansion of the US1-to-US3 region of recombinant viruses is diagrammed. The open box represents the internal junction between L and S components of the RhCMV genome. Cre/lox recombination was performed in Telo-RF cells. Recombinant clones containing the BAC vector in the viral genome (vRhCMV/BAC-EGFP) were plaque purified, and circular-form viral DNA was transformed into E. coli strain DH10B for plasmid isolation. (B) Substitution of the cre ORF for the egfp ORF by allelic exchange in E. coli . Only a portion of each plasmid is illustrated. The homologous flanking regions and the cre ORF of the recombination vector, pWC212, are illustrated. UL, unique sequences of the L component; US, unique sequences of the S component; PSV40, SV40 promoter; poly A, SV40 polyadenylation signal.

    Journal: Journal of Virology

    Article Title: Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis

    doi: 10.1128/JVI.77.9.5073-5083.2003

    Figure Lengend Snippet: Strategy for constructing a self-excisable RhCMV BAC. (A) Cloning of the full-length RhCMV genome into a BAC vector by Cre/lox recombination. The RhCMV genome structure with expansion of the US1-to-US3 region of recombinant viruses is diagrammed. The open box represents the internal junction between L and S components of the RhCMV genome. Cre/lox recombination was performed in Telo-RF cells. Recombinant clones containing the BAC vector in the viral genome (vRhCMV/BAC-EGFP) were plaque purified, and circular-form viral DNA was transformed into E. coli strain DH10B for plasmid isolation. (B) Substitution of the cre ORF for the egfp ORF by allelic exchange in E. coli . Only a portion of each plasmid is illustrated. The homologous flanking regions and the cre ORF of the recombination vector, pWC212, are illustrated. UL, unique sequences of the L component; US, unique sequences of the S component; PSV40, SV40 promoter; poly A, SV40 polyadenylation signal.

    Article Snippet: After incubation at 4°C for 24 h, cellular DNA and proteins were precipitated by centrifugation at 15,000 × g and 4°C for 30 min. DNA in the supernatant was extracted three times with phenol-chloroform, ethanol precipitated, and transformed into E. coli strain DH10B (Invitrogen) by electroporation according to published methods ( ).

    Techniques: BAC Assay, Clone Assay, Plasmid Preparation, Recombinant, Purification, Transformation Assay, Isolation

    Stability of the BAC pBACBHV37 over multiple generations in E. coli DH10B. DH10B cells containing pBACBHV37 were serially grown for 5, 10, and 20 days. BAC DNA was prepared from these cultures, and the restriction enzyme profiles were compared by FIGE with 1% agarose. DNA fragments were stained with ethidium bromide and photographed. Standard molecular sizes (in base pairs) are given.

    Journal: Journal of Virology

    Article Title: Construction and Manipulation of an Infectious Clone of the Bovine Herpesvirus 1 Genome Maintained as a Bacterial Artificial Chromosome

    doi: 10.1128/JVI.76.13.6660-6668.2002

    Figure Lengend Snippet: Stability of the BAC pBACBHV37 over multiple generations in E. coli DH10B. DH10B cells containing pBACBHV37 were serially grown for 5, 10, and 20 days. BAC DNA was prepared from these cultures, and the restriction enzyme profiles were compared by FIGE with 1% agarose. DNA fragments were stained with ethidium bromide and photographed. Standard molecular sizes (in base pairs) are given.

    Article Snippet: Aliquots of the ligation mixtures were electroporated into E. coli DH10B cells (1.5 kV, 100 Ω, 25 μF; Electroporator II; Invitrogen, San Diego, Calif.).

    Techniques: BAC Assay, Staining

    Drop assay of the four clones resistant to UV radiation to test perchlorate resistance. DH10B pBlueScript SK (+) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid medium supplemented with perchlorate 125 mM. An additional test was performed combining UVB (312 nm) radiation exposure during 80 s at 2.26 W m –2 and perchlorate (125 mM). A negative control without irradiation (–) was performed to verify the normal growth of the clones. Each experiment was performed at least three times using independent cultures.

    Journal: Frontiers in Microbiology

    Article Title: Novel Genes Involved in Resistance to Both Ultraviolet Radiation and Perchlorate From the Metagenomes of Hypersaline Environments

    doi: 10.3389/fmicb.2020.00453

    Figure Lengend Snippet: Drop assay of the four clones resistant to UV radiation to test perchlorate resistance. DH10B pBlueScript SK (+) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid medium supplemented with perchlorate 125 mM. An additional test was performed combining UVB (312 nm) radiation exposure during 80 s at 2.26 W m –2 and perchlorate (125 mM). A negative control without irradiation (–) was performed to verify the normal growth of the clones. Each experiment was performed at least three times using independent cultures.

    Article Snippet: Ligations were incubated overnight at 16°C using T4 DNA ligase (Roche) and were used to transform E. coli DH10B competent cells (Invitrogen) by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.

    Techniques: Clone Assay, Negative Control, Irradiation

    Drop assay of the four clones resistant to UV radiation and of the individual genes predicted in each clone. DH10B pBlueScript SK (+) (empty plasmid) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid and expose to radiation: (A) UVB (312 nm), 80 s at 2.26 W m –2 and (B) UVC (254 nm), 15 s at 1.56 W m –2 . Each experiment was performed at least three times using independent cultures.

    Journal: Frontiers in Microbiology

    Article Title: Novel Genes Involved in Resistance to Both Ultraviolet Radiation and Perchlorate From the Metagenomes of Hypersaline Environments

    doi: 10.3389/fmicb.2020.00453

    Figure Lengend Snippet: Drop assay of the four clones resistant to UV radiation and of the individual genes predicted in each clone. DH10B pBlueScript SK (+) (empty plasmid) was used as a negative control. 10 μl drops of serial dilutions of overnight cultures adjusted to an OD 600 = 1.0 were inoculated on LB solid and expose to radiation: (A) UVB (312 nm), 80 s at 2.26 W m –2 and (B) UVC (254 nm), 15 s at 1.56 W m –2 . Each experiment was performed at least three times using independent cultures.

    Article Snippet: Ligations were incubated overnight at 16°C using T4 DNA ligase (Roche) and were used to transform E. coli DH10B competent cells (Invitrogen) by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.

    Techniques: Clone Assay, Plasmid Preparation, Negative Control

    (A) Fluorescence microscopy images of DH10B cells expressing the hypothetical protein encoded by pML105- orf1 gene fused to GFP [DH10B pTU65- orf1 (105): gfp ]. On the left, cells growing exponentially before treatment with 4-NQO. On the right, cells from the same culture treated with 4-NQO (50 μM). White arrowheads indicate bright fluorescence foci. Below, microscopy images of selected cells are observed (GFP, DAPI staining and the overlay of both images). (B) Fluorescence microscopy images of DH10B cells expressing GFP constitutively [DH10B Δ rbsAR:cat_sf-gfp ]. On the left, cells growing exponentially before treatment with 4-NQO. On the right, cells from the same culture after treatment with 4-NQO (50 μM). Below, microscopy images of selected cells are observed (GFP, DAPI staining and the overlay of both images). White bars indicate 5 μm scale.

    Journal: Frontiers in Microbiology

    Article Title: Novel Genes Involved in Resistance to Both Ultraviolet Radiation and Perchlorate From the Metagenomes of Hypersaline Environments

    doi: 10.3389/fmicb.2020.00453

    Figure Lengend Snippet: (A) Fluorescence microscopy images of DH10B cells expressing the hypothetical protein encoded by pML105- orf1 gene fused to GFP [DH10B pTU65- orf1 (105): gfp ]. On the left, cells growing exponentially before treatment with 4-NQO. On the right, cells from the same culture treated with 4-NQO (50 μM). White arrowheads indicate bright fluorescence foci. Below, microscopy images of selected cells are observed (GFP, DAPI staining and the overlay of both images). (B) Fluorescence microscopy images of DH10B cells expressing GFP constitutively [DH10B Δ rbsAR:cat_sf-gfp ]. On the left, cells growing exponentially before treatment with 4-NQO. On the right, cells from the same culture after treatment with 4-NQO (50 μM). Below, microscopy images of selected cells are observed (GFP, DAPI staining and the overlay of both images). White bars indicate 5 μm scale.

    Article Snippet: Ligations were incubated overnight at 16°C using T4 DNA ligase (Roche) and were used to transform E. coli DH10B competent cells (Invitrogen) by electroporation using a Micropulser (Bio-Rad) according to the manufacturer’s instructions.

    Techniques: Fluorescence, Microscopy, Expressing, Staining

    A typical chimeric-GFP solubility screening experiment. ( A ) white light image, ( B ) transilluminating UV (365 nm) image of an agar plate used to select GFP-positive colonies in the solubility screen. Electrocompetent DH10B cells were transformed with a

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: A green fluorescent protein solubility screen in E. coli reveals domain boundaries of the GTP-binding domain in the P element transposase

    doi: 10.1002/pro.499

    Figure Lengend Snippet: A typical chimeric-GFP solubility screening experiment. ( A ) white light image, ( B ) transilluminating UV (365 nm) image of an agar plate used to select GFP-positive colonies in the solubility screen. Electrocompetent DH10B cells were transformed with a

    Article Snippet: 2–5 μL of each ligation was transformed into 40 μL electrocompetent DH10B cells (Invitrogen), diluted into 1 mL SOC medium (Invitrogen), then plated onto 5 selective agar plates.

    Techniques: Solubility, Transformation Assay

    Metalloid resistance conferred to Escherichia coli DH10B by metagenomic clone pA7 and pA12. Resistance to a Sodium arsenite, b Sodium arsenate and c Potassium antimony tartrate was examined with E. coli DH10B + pHB201(control) ( filled circle ), E. coli DH10B + pA7 ( filled triangle ) and E. coli DH10B + pA12 ( filled square ). Experiments performed in triplicates, values are averages and standard deviations are shown as error bars

    Journal: Indian Journal of Microbiology

    Article Title: Identification of Arsenic Resistance Genes from Marine Sediment Metagenome

    doi: 10.1007/s12088-017-0658-0

    Figure Lengend Snippet: Metalloid resistance conferred to Escherichia coli DH10B by metagenomic clone pA7 and pA12. Resistance to a Sodium arsenite, b Sodium arsenate and c Potassium antimony tartrate was examined with E. coli DH10B + pHB201(control) ( filled circle ), E. coli DH10B + pA7 ( filled triangle ) and E. coli DH10B + pA12 ( filled square ). Experiments performed in triplicates, values are averages and standard deviations are shown as error bars

    Article Snippet: The ligated product was electroporated at 200 Ω, 25 μf, and 12.5 kV/cm in E. coli DH10B using Micropulser II (Bio-Rad, USA).

    Techniques: