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    Promega pgem t easy vector
    Strategies for generation of dsRNAs and d-siRNAs. (A) <t>PCR</t> template strategy was utilized for obtaining s GFP , An rasA and An ras B dsRNA in a single T7 transcription reaction. T7 promoter sequence was added to both forward and reverse gene specific primers of s GFP , An rasA and An rasB , and then PCR amplification was performed in order to generate templates for dsRNA synthesis. (B) For generation of template DNA for unrelated MiAchE , PCR amplification was performed on pGEM T- MiAchE vector using M13 primers. This amplification includes the T7 promoter to one end and SP6 promoter to another end of MiAchE . (C) PCR templates for transcription of respective target genes. M. ladder; B. blank; s GFP , An rasA , An rasB and MiAchE target gene templates. (D) Synthesis of dsRNAs with T7 or SP6 in vitro transcription reactions. M. Ladder; s GFP , An rasA , An rasB and MiAchE dsRNAs. (E) 20% PAGE analysis of purified diced siRNAs. M-Ladder, s GFP , An rasA , An rasB and MiAchE d-siRNAs. The d-siRNAs of all target genes were generated by cleaving respective dsRNAs with RNase III at 37°C for 30 mins, followed by subsequent purifications and finally dissolved in nuclease free water.
    Pgem T Easy Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 95771 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Promega
    Average 99 stars, based on 95771 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-09
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    94
    Thermo Fisher pgem t easy vector
    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the <t>pGEM-T</t> vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).
    Pgem T Easy Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1311 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgem t easy vector/product/Thermo Fisher
    Average 94 stars, based on 1311 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

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    Strategies for generation of dsRNAs and d-siRNAs. (A) PCR template strategy was utilized for obtaining s GFP , An rasA and An ras B dsRNA in a single T7 transcription reaction. T7 promoter sequence was added to both forward and reverse gene specific primers of s GFP , An rasA and An rasB , and then PCR amplification was performed in order to generate templates for dsRNA synthesis. (B) For generation of template DNA for unrelated MiAchE , PCR amplification was performed on pGEM T- MiAchE vector using M13 primers. This amplification includes the T7 promoter to one end and SP6 promoter to another end of MiAchE . (C) PCR templates for transcription of respective target genes. M. ladder; B. blank; s GFP , An rasA , An rasB and MiAchE target gene templates. (D) Synthesis of dsRNAs with T7 or SP6 in vitro transcription reactions. M. Ladder; s GFP , An rasA , An rasB and MiAchE dsRNAs. (E) 20% PAGE analysis of purified diced siRNAs. M-Ladder, s GFP , An rasA , An rasB and MiAchE d-siRNAs. The d-siRNAs of all target genes were generated by cleaving respective dsRNAs with RNase III at 37°C for 30 mins, followed by subsequent purifications and finally dissolved in nuclease free water.

    Journal: PLoS ONE

    Article Title: Targeting Fungal Genes by Diced siRNAs: A Rapid Tool to Decipher Gene Function in Aspergillus nidulans

    doi: 10.1371/journal.pone.0075443

    Figure Lengend Snippet: Strategies for generation of dsRNAs and d-siRNAs. (A) PCR template strategy was utilized for obtaining s GFP , An rasA and An ras B dsRNA in a single T7 transcription reaction. T7 promoter sequence was added to both forward and reverse gene specific primers of s GFP , An rasA and An rasB , and then PCR amplification was performed in order to generate templates for dsRNA synthesis. (B) For generation of template DNA for unrelated MiAchE , PCR amplification was performed on pGEM T- MiAchE vector using M13 primers. This amplification includes the T7 promoter to one end and SP6 promoter to another end of MiAchE . (C) PCR templates for transcription of respective target genes. M. ladder; B. blank; s GFP , An rasA , An rasB and MiAchE target gene templates. (D) Synthesis of dsRNAs with T7 or SP6 in vitro transcription reactions. M. Ladder; s GFP , An rasA , An rasB and MiAchE dsRNAs. (E) 20% PAGE analysis of purified diced siRNAs. M-Ladder, s GFP , An rasA , An rasB and MiAchE d-siRNAs. The d-siRNAs of all target genes were generated by cleaving respective dsRNAs with RNase III at 37°C for 30 mins, followed by subsequent purifications and finally dissolved in nuclease free water.

    Article Snippet: The sGFP gene was PCR amplified from the pMT-sGFP vector and cloned in pGEM T-easy vector.

    Techniques: Polymerase Chain Reaction, Sequencing, Amplification, Plasmid Preparation, In Vitro, Polyacrylamide Gel Electrophoresis, Purification, Generated

    Expression of recombinant MAA2 in E. coli . (A) E. coli cells containing the maa2 gene ligated into vector pGEM-T in both orientations with respect to the lacZ promoter were grown in the presence and absence of IPTG, lysed, electrophoresed by SDS-PAGE on 7.5% (wt/vol) gels, transferred to nitrocellulose, and stained with MAA2-specific MAb 7a. Lanes contain M. arthritidis 158p10p9 whole-cell lysate (lane 1), E. coli containing the cloned insert in the forward orientation with respect to the lacZ promoter, in the absence (lane 2) and presence (lane 3) of IPTG, E. coli containing the cloned insert in the reverse orientation with respect to the lacZ promoter, in the absence (lane 4) and presence (lane 5) of IPTG, and untransformed E. coli in the absence (lane 6) and presence (lane 7) of IPTG. (B) E. coli cells containing the maa2 gene from M. arthritidis 158p10p9 clonal variants in which expression of MAA2 was switched ON or OFF were grown in the presence or absence of IPTG, lysed, electrophoresed and immunoblotted as described above. Lanes contain E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the reverse orientation with respect to the lacZ promoter, in the absence (lane 1) and presence (lane 2) of IPTG, E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 3) and presence (lane 4) of IPTG, E. coli in which maa2 from an ON variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 5) and presence (lane 6) of IPTG, and a whole-cell lysate of M. arthritidis strain 158p10p9 (lane 7).

    Journal: Infection and Immunity

    Article Title: Molecular Characterization of Mycoplasma arthritidis Variable Surface Protein MAA2

    doi:

    Figure Lengend Snippet: Expression of recombinant MAA2 in E. coli . (A) E. coli cells containing the maa2 gene ligated into vector pGEM-T in both orientations with respect to the lacZ promoter were grown in the presence and absence of IPTG, lysed, electrophoresed by SDS-PAGE on 7.5% (wt/vol) gels, transferred to nitrocellulose, and stained with MAA2-specific MAb 7a. Lanes contain M. arthritidis 158p10p9 whole-cell lysate (lane 1), E. coli containing the cloned insert in the forward orientation with respect to the lacZ promoter, in the absence (lane 2) and presence (lane 3) of IPTG, E. coli containing the cloned insert in the reverse orientation with respect to the lacZ promoter, in the absence (lane 4) and presence (lane 5) of IPTG, and untransformed E. coli in the absence (lane 6) and presence (lane 7) of IPTG. (B) E. coli cells containing the maa2 gene from M. arthritidis 158p10p9 clonal variants in which expression of MAA2 was switched ON or OFF were grown in the presence or absence of IPTG, lysed, electrophoresed and immunoblotted as described above. Lanes contain E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the reverse orientation with respect to the lacZ promoter, in the absence (lane 1) and presence (lane 2) of IPTG, E. coli in which maa2 from an OFF variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 3) and presence (lane 4) of IPTG, E. coli in which maa2 from an ON variant was ligated into vector pGEM-T in the forward orientation with respect to the lacZ promoter, in the absence (lane 5) and presence (lane 6) of IPTG, and a whole-cell lysate of M. arthritidis strain 158p10p9 (lane 7).

    Article Snippet: The full-length maa2 gene was amplified from strain H606 chromosomal DNA by PCR with primers F3 and HMPR and cloned into pGEM-T Easy as described above.

    Techniques: Expressing, Recombinant, Plasmid Preparation, SDS Page, Staining, Clone Assay, Variant Assay

    Agarose gel electrophoresis (1.2%) of the pGEM-T/CTRL-1 cDNA and pGEM-T/CTRL-1 genomic DNA after EcoR I digestion. M: 100-bp size marker; lane 1: full-length CTRL-1 cDNA PCR product; lane 2: pGEM-T/CTRL-1 cDNA; lane 3: CTRL-1 genomic DNA PCR product; lane 4: pGEM-T/CTRL-1 genomic DNA.

    Journal: Toxins

    Article Title: Cloning a Chymotrypsin-Like 1 (CTRL-1) Protease cDNA from the Jellyfish Nemopilema nomurai

    doi: 10.3390/toxins8070205

    Figure Lengend Snippet: Agarose gel electrophoresis (1.2%) of the pGEM-T/CTRL-1 cDNA and pGEM-T/CTRL-1 genomic DNA after EcoR I digestion. M: 100-bp size marker; lane 1: full-length CTRL-1 cDNA PCR product; lane 2: pGEM-T/CTRL-1 cDNA; lane 3: CTRL-1 genomic DNA PCR product; lane 4: pGEM-T/CTRL-1 genomic DNA.

    Article Snippet: All the PCR products were purified with the Expin™ GeneAll® PCR SV purification kit (GeneAll, Seoul, Korea), cloned into the pGEM®-T Easy Vector System (Promega, Madison, WI, USA) and confirmed the clone by EcoR I digestion at 37 °C for 1 h. Full-length of cDNA sequence was identified by ABI PRISM 3739 Genetic Analyzer (Thermo Fisher, Waltham, MA, USA).

    Techniques: Agarose Gel Electrophoresis, Marker, Polymerase Chain Reaction

    Analysis of electrophoretic profiles. a Agarose gel profile of products resulting from PCR amplification of the VEGF transcripts present in tumor tissue: human breast cancer cell line MCF7 ( lane 2 ), human colorectal cancer cells ( lane 4 ) and human breast cancer cells ( lane 5 ) and in distant-tumor tissue ( lane 3 ) M molecular-mass marker (100 pb DNA ladder; Fermentas); lane 1 PCR negative control (sample without DNA). b Agarose gel showing PCR products from amplification of the positive clones obtained after ligation to pGEMT easy vector. Lane 1 VEGF 121 , lane 2/3 VEGF 165 , lane 4/5 VEGF 189 , T-: PCR negative control (sample without DNA)

    Journal: AMB Express

    Article Title: Expression, purification and functionality of bioactive recombinant human vascular endothelial growth factor VEGF165 in E. coli

    doi: 10.1186/s13568-016-0300-2

    Figure Lengend Snippet: Analysis of electrophoretic profiles. a Agarose gel profile of products resulting from PCR amplification of the VEGF transcripts present in tumor tissue: human breast cancer cell line MCF7 ( lane 2 ), human colorectal cancer cells ( lane 4 ) and human breast cancer cells ( lane 5 ) and in distant-tumor tissue ( lane 3 ) M molecular-mass marker (100 pb DNA ladder; Fermentas); lane 1 PCR negative control (sample without DNA). b Agarose gel showing PCR products from amplification of the positive clones obtained after ligation to pGEMT easy vector. Lane 1 VEGF 121 , lane 2/3 VEGF 165 , lane 4/5 VEGF 189 , T-: PCR negative control (sample without DNA)

    Article Snippet: The PCR product mixture obtained after amplification of the MCF7 cDNA was ligated into pGEMT easy vector and cloned in E. coli Top10.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Amplification, Marker, Negative Control, Clone Assay, Ligation, Plasmid Preparation

    pELMO and pGEM-T Easy vector cloning efficiency for different sized inserts. pELMO and pGEM-T Easy cloning efficiency regarding low, medium and large sized inserts. Cloning efficiency is expressed as the ratio of the amount of PCR positive colonies

    Journal: AMB Express

    Article Title: pELMO, an optimised in-house cloning vector

    doi: 10.1186/s13568-017-0324-2

    Figure Lengend Snippet: pELMO and pGEM-T Easy vector cloning efficiency for different sized inserts. pELMO and pGEM-T Easy cloning efficiency regarding low, medium and large sized inserts. Cloning efficiency is expressed as the ratio of the amount of PCR positive colonies

    Article Snippet: Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Techniques: Plasmid Preparation, Clone Assay, Polymerase Chain Reaction

    Construction of full-length cDNA clone of VHSV DK-3952B genome. Six overlapping cDNA fragments covering the entire VHSV genome were assembled by ligation into a pCI vector using the EcoRI, PvuII, EcoRV, KpnI, SalI, BclI, SmaI, StuI, BsrGI and NotI restriction sites. Assembly was carried out in 3 steps; (i) cloning of six fragments (F1-F6) separately in pGEM-T vectors (see methods), and addition of hammerhead ribozyme (HHRz) at the 5′-end of F1 and hepatitis delta ribozyme (HdvRz) at the 3′-end of F6; (ii) construction of A, B and C clones into the pCI vector; (iii) assembly of the full-length clone by ligating fragments from these three clones in the pCI vector. Restriction sites artificially created (*) and naturally present in the genome are indicated. Abbreviation: CMV, cytomegalovirus immediate-early enhancer and promoter

    Journal: Virology Journal

    Article Title: The glycoprotein, non-virion protein, and polymerase of viral hemorrhagic septicemia virus are not determinants of host-specific virulence in rainbow trout

    doi: 10.1186/s12985-019-1139-3

    Figure Lengend Snippet: Construction of full-length cDNA clone of VHSV DK-3952B genome. Six overlapping cDNA fragments covering the entire VHSV genome were assembled by ligation into a pCI vector using the EcoRI, PvuII, EcoRV, KpnI, SalI, BclI, SmaI, StuI, BsrGI and NotI restriction sites. Assembly was carried out in 3 steps; (i) cloning of six fragments (F1-F6) separately in pGEM-T vectors (see methods), and addition of hammerhead ribozyme (HHRz) at the 5′-end of F1 and hepatitis delta ribozyme (HdvRz) at the 3′-end of F6; (ii) construction of A, B and C clones into the pCI vector; (iii) assembly of the full-length clone by ligating fragments from these three clones in the pCI vector. Restriction sites artificially created (*) and naturally present in the genome are indicated. Abbreviation: CMV, cytomegalovirus immediate-early enhancer and promoter

    Article Snippet: The cDNA fragments obtained after RT-PCR amplification were cloned into pGEM-T or pGEM-T Easy vectors (Promega Corp., Madison, WI).

    Techniques: Ligation, Plasmid Preparation, Clone Assay

    Amplification of the housekeeping genes and confirmation of cloning by EcoRI digestion of pGEM-T vector. (A) Amplification of the six target genes from cDNA of eggplant Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: GAPDH (586bp), 4: Cyclophilin (265 bp), 5: Actin (333 bp), 6: RuBP (269bp). (B) Confirmation of the inserts in the recombinant pGEM-T vector by EcoRI digestion. Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: Cyclophilin (265 bp), 4: GAPDH (586bp), 5: Actin (333 bp), 6: RuBP (269bp). (C) Amplification of the six target genes from genomic DNA of eggplant. Lanes- M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (No amplification), 3: Cyclophilin (265 bp), 4: GAPDH (1.2 kb), 5: Actin (333 bp), 6: RuBP (269bp).

    Journal: BMC Research Notes

    Article Title: Selection and validation of reference genes for quantitative gene expression studies by real-time PCR in eggplant (Solanum melongena L)

    doi: 10.1186/1756-0500-6-312

    Figure Lengend Snippet: Amplification of the housekeeping genes and confirmation of cloning by EcoRI digestion of pGEM-T vector. (A) Amplification of the six target genes from cDNA of eggplant Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: GAPDH (586bp), 4: Cyclophilin (265 bp), 5: Actin (333 bp), 6: RuBP (269bp). (B) Confirmation of the inserts in the recombinant pGEM-T vector by EcoRI digestion. Lanes - M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (454 bp), 3: Cyclophilin (265 bp), 4: GAPDH (586bp), 5: Actin (333 bp), 6: RuBP (269bp). (C) Amplification of the six target genes from genomic DNA of eggplant. Lanes- M: 100 bp DNA Ladder/Marker, 1. 18S rRNA (416bp), 2: APRT (No amplification), 3: Cyclophilin (265 bp), 4: GAPDH (1.2 kb), 5: Actin (333 bp), 6: RuBP (269bp).

    Article Snippet: The amplified cDNA PCR products were cloned into pGEM-T Easy cloning vector (Promega) according to the manufacturer’s instructions.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Marker, Recombinant

    Transformation frequencies of H. pylori mutants. (A) H. pylori strain 26695 and derivative strains were transformed by homologous recombination with p801R (an 801-bp H. pylori rpsL fragment from a streptomycin-resistant [Str r ] strain cloned into pGEM-T

    Journal: Journal of Bacteriology

    Article Title: DprB Facilitates Inter- and Intragenomic Recombination in Helicobacter pylori

    doi: 10.1128/JB.00346-12

    Figure Lengend Snippet: Transformation frequencies of H. pylori mutants. (A) H. pylori strain 26695 and derivative strains were transformed by homologous recombination with p801R (an 801-bp H. pylori rpsL fragment from a streptomycin-resistant [Str r ] strain cloned into pGEM-T

    Article Snippet: An 801-bp DNA sequence encompassing H. pylori rpsL from Strr H. pylori strain 26695 ( , ) was obtained by PCR using primers rpsL801-F and rpsL801-R (see Table S2 in the supplemental material) and cloned into the vector pGEM-T Easy (Promega, Madison, WI) to create plasmid p801R (see Table S1), which was used as donor DNA for homologous transformation.

    Techniques: Transformation Assay, Homologous Recombination, Clone Assay

    Cross-species complementation. (A) Complementation of the E. coli ruvC mutant by the cloned H. pylori dprB and ruvC genes. E. coli cells carrying pGEM-T Easy-derived plasmids on an LB plate with ampicillin were subjected to a range of UV exposures, and

    Journal: Journal of Bacteriology

    Article Title: DprB Facilitates Inter- and Intragenomic Recombination in Helicobacter pylori

    doi: 10.1128/JB.00346-12

    Figure Lengend Snippet: Cross-species complementation. (A) Complementation of the E. coli ruvC mutant by the cloned H. pylori dprB and ruvC genes. E. coli cells carrying pGEM-T Easy-derived plasmids on an LB plate with ampicillin were subjected to a range of UV exposures, and

    Article Snippet: An 801-bp DNA sequence encompassing H. pylori rpsL from Strr H. pylori strain 26695 ( , ) was obtained by PCR using primers rpsL801-F and rpsL801-R (see Table S2 in the supplemental material) and cloned into the vector pGEM-T Easy (Promega, Madison, WI) to create plasmid p801R (see Table S1), which was used as donor DNA for homologous transformation.

    Techniques: Mutagenesis, Clone Assay, Derivative Assay

    Schematic representation of the capsule genes of W135 serogroup in transcriptional fusion of synG with ermAM cassette . The synG gene responsible for the synthesis of the W135 capsule was amplified using the 98-30 and 03-12-5 oligonucleotides (Table 3) from W135 ATCC strain. The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11. In the same conditions, another fragment was amplified using the 04.02-2/galECK29A from synG downstream sequence to generate pLAN52. The ermAM cassette was insered into Nco I site of pLAN52 to generate pLAN53. The fragment amplified from pLAN53 with the ERAM1 and galECK29A (Dolan Livengood [ 23 ] et al ., 2003) was insered into Pst I site of pLAN11 to generate pLAN13-2. This plasmid was linearised by the enzyme SphI and transformed into W135 ATCC strain to generate the synG::ermAM strain M6, erythromycin resistant.

    Journal: Journal of Nanobiotechnology

    Article Title: Effect of mesoporous silica under Neisseria meningitidis transformation process: environmental effects under meningococci transformation

    doi: 10.1186/1477-3155-9-28

    Figure Lengend Snippet: Schematic representation of the capsule genes of W135 serogroup in transcriptional fusion of synG with ermAM cassette . The synG gene responsible for the synthesis of the W135 capsule was amplified using the 98-30 and 03-12-5 oligonucleotides (Table 3) from W135 ATCC strain. The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11. In the same conditions, another fragment was amplified using the 04.02-2/galECK29A from synG downstream sequence to generate pLAN52. The ermAM cassette was insered into Nco I site of pLAN52 to generate pLAN53. The fragment amplified from pLAN53 with the ERAM1 and galECK29A (Dolan Livengood [ 23 ] et al ., 2003) was insered into Pst I site of pLAN11 to generate pLAN13-2. This plasmid was linearised by the enzyme SphI and transformed into W135 ATCC strain to generate the synG::ermAM strain M6, erythromycin resistant.

    Article Snippet: The amplified fragment was cloned into the pGEM-T Easy Vector System I (Promega, Madison, WI, USA), to generate the plasmid pLAN11.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Sequencing, Transformation Assay

    Schematic representation of the capsule genes of C serogroup in disrupted construction of NMB0065 gene with aaDA cassette . The NMB0065 gene was amplified using the 03-12-3 and 03-12-4 oligonucleotides (Table 3) from C2135 strain. This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6. E. coli strain Z501 was transformed with plasmid pLAN6 resulting in the plasmid pLAN7. The Ω aaDA cassette was inserted into the BclI site of pLAN7 to generate plasmid pLAN45, which was transformed into the C2135 strain to generate the isogenic mutant strain M2.

    Journal: Journal of Nanobiotechnology

    Article Title: Effect of mesoporous silica under Neisseria meningitidis transformation process: environmental effects under meningococci transformation

    doi: 10.1186/1477-3155-9-28

    Figure Lengend Snippet: Schematic representation of the capsule genes of C serogroup in disrupted construction of NMB0065 gene with aaDA cassette . The NMB0065 gene was amplified using the 03-12-3 and 03-12-4 oligonucleotides (Table 3) from C2135 strain. This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6. E. coli strain Z501 was transformed with plasmid pLAN6 resulting in the plasmid pLAN7. The Ω aaDA cassette was inserted into the BclI site of pLAN7 to generate plasmid pLAN45, which was transformed into the C2135 strain to generate the isogenic mutant strain M2.

    Article Snippet: This fragment was cloned into the pGEM-T Easy Vector System II (Promega Corporation, Madison, WI, USA), to generate the plasmid pLAN6.

    Techniques: Amplification, Clone Assay, Plasmid Preparation, Transformation Assay, Mutagenesis

    Export of constitutively expressed immunity protein. E. coli DH5α cells containing the ximB locus with an 5′-upstream promoter sequence in pGEM-T Easy vector was grown overnight. Cytosolic, periplasmic, and flagellar proteins were prepared

    Journal: Journal of Bacteriology

    Article Title: Xenocin Export by the Flagellar Type III Pathway in Xenorhabdus nematophila

    doi: 10.1128/JB.01532-12

    Figure Lengend Snippet: Export of constitutively expressed immunity protein. E. coli DH5α cells containing the ximB locus with an 5′-upstream promoter sequence in pGEM-T Easy vector was grown overnight. Cytosolic, periplasmic, and flagellar proteins were prepared

    Article Snippet: The plasmid vector pGEM-T Easy, from Promega (Madison, WI), was used for PCR cloning. pCR-XL-TOPO and pBCSK(+) were used for cloning and complementation studies.

    Techniques: Sequencing, Plasmid Preparation

    DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the pGEM-T vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).

    Journal: Molecular Vision

    Article Title: FGFR2 molecular analysis and related clinical findings in one Chinese family with Crouzon Syndrome

    doi:

    Figure Lengend Snippet: DNA sequence of a part of FGF2 in the affected patients and unaffected individuals. A : A mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. B : Sequence of the normal allele of exon 8 subcloned into the pGEM-T vector used as a control. C : A heterozygous missense mutation c.866A > C (Gln289Pro) in exon 8 in the affected individuals. The mutation causes the glutarnine 289 codon (CAG) to change to a proline codon (CCG).

    Article Snippet: The purified PCR fragments were ligated into the pGEM-T easy vector (Invitrogen, Carlsbad, CA), and the resulting plasmids were transfected, by heat shock, into DH5a-competent Escherichia coli for propagation.

    Techniques: Sequencing, Mutagenesis, Plasmid Preparation