pgem t easy vector promega  (Promega)

 
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    Name:
    pGEM T Easy Vector Systems
    Description:
    PCR cloning vectors with 3 options for insert excision
    Catalog Number:
    a1360
    Price:
    None
    Category:
    Nucleic Acid Extraction Analysis PCR PCR Cloning
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    Structured Review

    Promega pgem t easy vector promega
    PCR cloning vectors with 3 options for insert excision
    https://www.bioz.com/result/pgem t easy vector promega/product/Promega
    Average 90 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    pgem t easy vector promega - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Methylation Sequencing:

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: Paragraph title: Genomic DNA preparation and bisulfite sequencing analysis ... Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing.

    Clone Assay:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence. .. Orthologs of acdS and acdR were retrieved from GenBank database running a blast search over Rhizobiaceae (taxid:82115) non-redundant nucleotide database on 2016-05-16 by using acdS gene from AK83 (Sinme_5642) and acdR from AK83 (Sinme_5643) as query sequence.

    Article Title: Type-I Prenyl Protease Function Is Required in the Male Germline of Drosophila melanogaster
    Article Snippet: .. The cDNAs were amplified with a high-fidelity Taq (Phusion M05030S) and cloned into the EcoRI sites of pGEM (Promega PR-A1360). ..

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: .. The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers. .. The Cytochrome c oxidase subunite 1 (Cox1) gene fragment was amplified using JB3 ( TTT TTT GGG CAT CCT GAG GTT TAT ) and JB4.5 ( TAA AGA AAG AAC ATA ATG AAA ATG ) [ , ].

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: .. The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA). .. The plasmids were sequenced using ABI 3130 DNA sequencer (Applied Biosystems, California, USA) and using three primers (T7, SP6m and Ura3_R2).

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: The pELMO vector (3306 bp) was constructed for the efficient and reliable cloning of PCR products; it contains two selection systems: the most common ampicillin-resistance marker generally used in basic research groups and the ccdB gene encoding a 101 amino acid toxic protein expressed by the lac promoter. .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Recombinant clones were sequenced with an automatic DNA sequencer (ABI, USA) and spliced artificially.

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: .. ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions. .. Inserts of 6–18 positive clones (depending on the ploidy level: 6 clones per diploid genome) were amplified using colony-PCR with universal M13 primers whereby recombinant colonies were added directly into the PCR mastermix and inserts amplified using reagents and conditions described in .

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). ..

    Centrifugation:

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: After centrifugation and separation, the aqueous phase was used to purify total RNA by RNeasy Mini kit (Qiagen GmBH, Germany) following the manufacturer’s protocol. .. The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA).

    Amplification:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence. .. Orthologs of acdS and acdR were retrieved from GenBank database running a blast search over Rhizobiaceae (taxid:82115) non-redundant nucleotide database on 2016-05-16 by using acdS gene from AK83 (Sinme_5642) and acdR from AK83 (Sinme_5643) as query sequence.

    Article Title: Type-I Prenyl Protease Function Is Required in the Male Germline of Drosophila melanogaster
    Article Snippet: .. The cDNAs were amplified with a high-fidelity Taq (Phusion M05030S) and cloned into the EcoRI sites of pGEM (Promega PR-A1360). ..

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: .. The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers. .. The Cytochrome c oxidase subunite 1 (Cox1) gene fragment was amplified using JB3 ( TTT TTT GGG CAT CCT GAG GTT TAT ) and JB4.5 ( TAA AGA AAG AAC ATA ATG AAA ATG ) [ , ].

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: The URA3 cDNA was amplified by PCR using semi-nested PCR with three primers (1st , Ura3c_F1 and Ura3c_R1; 2nd , Ura3c_F1 and Ura3c_R2) (Table ) using Tks Gflex DNA polymerase (Takara Bio, Japan). .. The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA).

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: Reverse transcription PCR (RT-PCR) was performed using SuperScript III reverse transcriptase (Invitrogen, USA) for reverse transcription and PfuUltra high-fidelity DNA polymerase (Stratagene, USA) for PCR amplification. .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA).

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: .. Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing. .. The primer sets used for the bisulfite sequencing are as follows: 5′-TGGGTTGAAATATTGGGTTTATTT-3′and 5′-CTAAA AC CAAATATCCAACCATA-3′ for the Oct4 gene, 5′-GATTTGTAG GTGGGATTAATTGTGAATTT-3′and 5′-ACCAAAAA AACCCA CACTC ATATCAATATA-3′ for the Nanog gene ( ).

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions. .. Inserts of 6–18 positive clones (depending on the ploidy level: 6 clones per diploid genome) were amplified using colony-PCR with universal M13 primers whereby recombinant colonies were added directly into the PCR mastermix and inserts amplified using reagents and conditions described in .

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). ..

    Article Title: The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1 [OPEN]
    Article Snippet: .. The fragment amplified was then introduced into the pGEM vector by pGEM-T (Easy) Vector Systems (Promega). .. To generate standard curves for absolute quantification, a series of dilutions (from 1 × 10−1 to 10−6 ng) of the plasmids prepared were made and then assayed by real-time PCR.

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Synthesized:

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). .. Linearized plasmid, extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), was used as a template and antisense RNA probe was synthesized using T7 enzyme mix (AM1320, MaxiScript SP6/T7 In Vitro Transcription Kit, Ambion, USA) and DIG-labelling mix (11277073910, DIG RNA Labelling Mix, Roche, Germany).

    Construct:

    Article Title: Type-I Prenyl Protease Function Is Required in the Male Germline of Drosophila melanogaster
    Article Snippet: Paragraph title: Rescue constructs: ... The cDNAs were amplified with a high-fidelity Taq (Phusion M05030S) and cloned into the EcoRI sites of pGEM (Promega PR-A1360).

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: This latter mechanism allows direct selection of positive recombinants by disrupting lethal genes, as shown in similarly constructed vectors (Bernard ; Gabant et al. ). ccdB expression thus results in the death of cells containing a non-recombinant vector, offering a highly efficient, positive selection system, even being comparable with white/blue selection systems based on the LacZ operon, one of the most used for this purpose (Bernard ; Messing et al. ). .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1 [OPEN]
    Article Snippet: The fragment amplified was then introduced into the pGEM vector by pGEM-T (Easy) Vector Systems (Promega). .. To generate standard curves for absolute quantification, a series of dilutions (from 1 × 10−1 to 10−6 ng) of the plasmids prepared were made and then assayed by real-time PCR.

    Incubation:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. Following overnight incubation at 37°C, white colonies were picked, cultured in ampicillin-enriched LB medium, and amplified.

    TALENs:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: Paragraph title: Generation of TALENs-targeted Hnrnph1 +/- and Rufy1 +/- mice ... To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    Expressing:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: This latter mechanism allows direct selection of positive recombinants by disrupting lethal genes, as shown in similarly constructed vectors (Bernard ; Gabant et al. ). ccdB expression thus results in the death of cells containing a non-recombinant vector, offering a highly efficient, positive selection system, even being comparable with white/blue selection systems based on the LacZ operon, one of the most used for this purpose (Bernard ; Messing et al. ). .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1 [OPEN]
    Article Snippet: Paragraph title: Absolute Expression Analysis of Nramp5 in Barley and Rice ... The fragment amplified was then introduced into the pGEM vector by pGEM-T (Easy) Vector Systems (Promega).

    Transformation Assay:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA). .. This strain lacks the F plasmid which encodes the CCDA protein; this product acts as inhibitor of ccdB function (Van Melderen et al. ). pELMO vector transformation efficiency was ascertained by cloning csp , msp1 and eba -175 PCR products through blunt-end ligation into pELMO.

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Ligation:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA). .. This strain lacks the F plasmid which encodes the CCDA protein; this product acts as inhibitor of ccdB function (Van Melderen et al. ). pELMO vector transformation efficiency was ascertained by cloning csp , msp1 and eba -175 PCR products through blunt-end ligation into pELMO.

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Cell Culture:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. Following overnight incubation at 37°C, white colonies were picked, cultured in ampicillin-enriched LB medium, and amplified.

    Generated:

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Polymerase Chain Reaction:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: Detection of acdS gene, genomic context, analysis, and phylogenetic reconstruction The presence of acdS orthologs in a collection of 133 S. meliloti strains was performed by PCR amplification on crude lysates using the two sets of primers and the PCR conditions described in Duan et al. ( ). .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence.

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: Paragraph title: DNA extraction, PCR amplification and sequencing ... The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers.

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: .. The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA). .. The plasmids were sequenced using ABI 3130 DNA sequencer (Applied Biosystems, California, USA) and using three primers (T7, SP6m and Ura3_R2).

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: A primer set was designed (ccdBSec-Dir/ccdBSec-Rev) for sequencing plasmid inserts. pELMO digestion with the Sma I enzyme produced a blunt-ended vector; this bypassed the A-tailing reaction step for PCR fragments obtained by high fidelity polymerases. .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: Reverse transcription PCR (RT-PCR) was performed using SuperScript III reverse transcriptase (Invitrogen, USA) for reverse transcription and PfuUltra high-fidelity DNA polymerase (Stratagene, USA) for PCR amplification. .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA).

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: The cycle sequencing reactions were performed using the same primers as for the PCR amplifications and internal primers where appropriate ( ). .. ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions.

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: The region between primer pair 5 (forward) and primer pair 6 (reverse) from Table , which is the corresponding region to the mutation site in patients, was amplified from 24 hpf embryo cDNA under the following PCR conditions: 2 min at 95 °C, 30 s at 95 °C, 30 s at 62 °C, 2 min at 72 °C for 35 cycles; and 7 min at 72 °C. .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA).

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Sequencing:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence. .. Orthologs of acdS and acdR were retrieved from GenBank database running a blast search over Rhizobiaceae (taxid:82115) non-redundant nucleotide database on 2016-05-16 by using acdS gene from AK83 (Sinme_5642) and acdR from AK83 (Sinme_5643) as query sequence.

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: Paragraph title: DNA extraction, PCR amplification and sequencing ... The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers.

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: Paragraph title: RNA extraction and determining the URA3 transcript sequence ... The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA).

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: A primer set was designed (ccdBSec-Dir/ccdBSec-Rev) for sequencing plasmid inserts. pELMO digestion with the Sma I enzyme produced a blunt-ended vector; this bypassed the A-tailing reaction step for PCR fragments obtained by high fidelity polymerases. .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: Paragraph title: RNA isolation and genome sequencing. ... The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA).

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: .. Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing. .. The primer sets used for the bisulfite sequencing are as follows: 5′-TGGGTTGAAATATTGGGTTTATTT-3′and 5′-CTAAA AC CAAATATCCAACCATA-3′ for the Oct4 gene, 5′-GATTTGTAG GTGGGATTAATTGTGAATTT-3′and 5′-ACCAAAAA AACCCA CACTC ATATCAATATA-3′ for the Nanog gene ( ).

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: Sequencing reactions were run on a 3130xl Genetic Analyzer automated capillary sequencer (Applied Biosystems). .. ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions.

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Injection:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: Each mRNA cocktail was diluted in sterile buffer and injected into B6 single-cell embryos at the BUMC Transgenic Core facility ( http://www.bumc.bu.edu/transgenic/ ). .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    Recombinant:

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Recombinant clones were sequenced with an automatic DNA sequencer (ABI, USA) and spliced artificially.

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions. .. Inserts of 6–18 positive clones (depending on the ploidy level: 6 clones per diploid genome) were amplified using colony-PCR with universal M13 primers whereby recombinant colonies were added directly into the PCR mastermix and inserts amplified using reagents and conditions described in .

    Cellular Antioxidant Activity Assay:

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: The 18S rDNA gene was amplified using G18S4 ( GCT TGT CTC AAA GAT TAA GCC ) and 18P ( TGA TCC WKC YGC AGG TTC AC ) with internal sequencing primers 4F ( CAA GGA CGA WAG TTW GAG G ) and 4R ( GTA TCT GAT CGC CKT CGA WC ) according to Bert et al. [ ]. .. The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers.

    DNA Extraction:

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: Paragraph title: DNA extraction, PCR amplification and sequencing ... The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers.

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: Genomic DNA preparation and bisulfite sequencing analysis Genomic DNA of each cell line was extracted with a Genomic DNA extraction kit (17045)(Intron Biotechnology, korea). .. Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing.

    Mutagenesis:

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: The region between primer pair 5 (forward) and primer pair 6 (reverse) from Table , which is the corresponding region to the mutation site in patients, was amplified from 24 hpf embryo cDNA under the following PCR conditions: 2 min at 95 °C, 30 s at 95 °C, 30 s at 62 °C, 2 min at 72 °C for 35 cycles; and 7 min at 72 °C. .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA).

    Isolation:

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: Paragraph title: RNA isolation and genome sequencing. ... The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA).

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Negative Control:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: S. meliloti 1021 was used as negative control, while S. meliloti AK83 was used as positive controls. .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence.

    Purification:

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers. .. Sanger sequencing of purified PCR fragments was carried out in forward and reverse direction by Macrogen (Europe).

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Recombinant clones were sequenced with an automatic DNA sequencer (ABI, USA) and spliced artificially.

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: .. Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing. .. The primer sets used for the bisulfite sequencing are as follows: 5′-TGGGTTGAAATATTGGGTTTATTT-3′and 5′-CTAAA AC CAAATATCCAACCATA-3′ for the Oct4 gene, 5′-GATTTGTAG GTGGGATTAATTGTGAATTT-3′and 5′-ACCAAAAA AACCCA CACTC ATATCAATATA-3′ for the Nanog gene ( ).

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: The purified fragments were directly sequenced using dye terminator chemistry following the manufacturer’s protocol (Applied Biosystems). .. ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions.

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The PCR product was purified using the QIAprep Miniprep kit (QIAGEN).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: Reverse transcription PCR (RT-PCR) was performed using SuperScript III reverse transcriptase (Invitrogen, USA) for reverse transcription and PfuUltra high-fidelity DNA polymerase (Stratagene, USA) for PCR amplification. .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA).

    Selection:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: This latter mechanism allows direct selection of positive recombinants by disrupting lethal genes, as shown in similarly constructed vectors (Bernard ; Gabant et al. ). ccdB expression thus results in the death of cells containing a non-recombinant vector, offering a highly efficient, positive selection system, even being comparable with white/blue selection systems based on the LacZ operon, one of the most used for this purpose (Bernard ; Messing et al. ). .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Staining:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: Agarose gel electrophoresis on 1.5% TAE buffer and ethidium bromide staining (10 mg/l) was used for visualization of amplification products on an UV transilluminator. .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers. .. The Cytochrome c oxidase subunite 1 (Cox1) gene fragment was amplified using JB3 ( TTT TTT GGG CAT CCT GAG GTT TAT ) and JB4.5 ( TAA AGA AAG AAC ATA ATG AAA ATG ) [ , ].

    Mouse Assay:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: Paragraph title: Generation of TALENs-targeted Hnrnph1 +/- and Rufy1 +/- mice ... To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    In Situ Hybridization:

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: Paragraph title: Whole mount in situ hybridization (WMISH) ... The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA).

    Plasmid Preparation:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence. .. Orthologs of acdS and acdR were retrieved from GenBank database running a blast search over Rhizobiaceae (taxid:82115) non-redundant nucleotide database on 2016-05-16 by using acdS gene from AK83 (Sinme_5642) and acdR from AK83 (Sinme_5643) as query sequence.

    Article Title: Integrative taxonomy of root-knot nematodes reveals multiple independent origins of mitotic parthenogenesis
    Article Snippet: .. The internal transcribed rDNA spacer (ITS) was amplified using VRAIN2F ( CTT TGT ACA CAC CGC CCG TCG CT ) and VRAIN2R ( TTT CAC TCG CCG TTA CTA AGG GAA TC ) subsequently cloned using pGEM ® -T easy vector systems (Promega) and sequenced using universal M13F and M13R primers. .. The Cytochrome c oxidase subunite 1 (Cox1) gene fragment was amplified using JB3 ( TTT TTT GGG CAT CCT GAG GTT TAT ) and JB4.5 ( TAA AGA AAG AAC ATA ATG AAA ATG ) [ , ].

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: .. The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA). .. The plasmids were sequenced using ABI 3130 DNA sequencer (Applied Biosystems, California, USA) and using three primers (T7, SP6m and Ura3_R2).

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA). .. The results indicated that pELMO is suitable for cloning in the E. coli TOP10 strain as it does not carry the lacI q repressor, therefore granting constitutive expression of ccdB product without the need for IPTG (isopropyl-β-d -thiogalactopyranoside) induction.

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Recombinant clones were sequenced with an automatic DNA sequencer (ABI, USA) and spliced artificially.

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: .. Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Imprinted gene Zinc finger protein 127 is a novel regulator of master pluripotency transcription factor, Oct4
    Article Snippet: .. Amplified DNA was then purified and ligated to the pGEM-T easy vector (A1360)(Promega, USA) for sequencing. .. The primer sets used for the bisulfite sequencing are as follows: 5′-TGGGTTGAAATATTGGGTTTATTT-3′and 5′-CTAAA AC CAAATATCCAACCATA-3′ for the Oct4 gene, 5′-GATTTGTAG GTGGGATTAATTGTGAATTT-3′and 5′-ACCAAAAA AACCCA CACTC ATATCAATATA-3′ for the Nanog gene ( ).

    Article Title: Molecular phylogenetic analyses of nuclear and plastid DNA sequences support dysploid and polyploid chromosome number changes and reticulate evolution in the diversification of Melampodium (Millerieae, Asteraceae)
    Article Snippet: .. ITS sequences of diploid accessions that showed double/multiple peaks, as well as of all polyploid accessions, were cloned using the pGEM-T-easy vector systems and JM109 competent cells (Promega, Madison, WI, USA) following manufacturer’s instructions. .. Inserts of 6–18 positive clones (depending on the ploidy level: 6 clones per diploid genome) were amplified using colony-PCR with universal M13 primers whereby recombinant colonies were added directly into the PCR mastermix and inserts amplified using reagents and conditions described in .

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). ..

    Article Title: The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1The HvNramp5 Transporter Mediates Uptake of Cadmium and Manganese, But Not Iron 1 [OPEN]
    Article Snippet: .. The fragment amplified was then introduced into the pGEM vector by pGEM-T (Easy) Vector Systems (Promega). .. To generate standard curves for absolute quantification, a series of dilutions (from 1 × 10−1 to 10−6 ng) of the plasmids prepared were made and then assayed by real-time PCR.

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega). .. The ligation reaction was transformed into MAX Efficiency DH5α Competent Cells (Invitrogen) and plated onto Ampicillin-IPTG/X-Gal LB agarose plates for blue-white selection.

    Software:

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Clustal X 1.83 and DNAStar software were used to analyze the sequences.

    RNA Extraction:

    Article Title: Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies
    Article Snippet: Paragraph title: RNA extraction and determining the URA3 transcript sequence ... The product of the second PCR was tailed with adenine using Taq polymerase (Takara Bio, Japan) and cloned into pGEM-T Easy Vector Systems (Promega, Madison, USA).

    Binding Assay:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: Genomic DNA was extracted from mouse tail biopsies and PCR-amplified with primers targeting100 base pairs upstream and downstream of the TALENs binding domain. .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    Agarose Gel Electrophoresis:

    Article Title: Role and Regulation of ACC Deaminase Gene in Sinorhizobium meliloti: Is It a Symbiotic, Rhizospheric or Endophytic Gene?
    Article Snippet: Agarose gel electrophoresis on 1.5% TAE buffer and ethidium bromide staining (10 mg/l) was used for visualization of amplification products on an UV transilluminator. .. Positive amplification products from two strains, representative of the collection (BO21CC, 2B13) were cloned into pGEM®-T Easy Vector Systems (Promega) following manufacturer's instructions and sequenced for confirmation of acdS presence.

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: .. The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). ..

    In Vitro:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: Several positive selection system-based vectors have been considered efficient tools to date for simplifying in vitro DNA recombination procedures (Liu et al. ). .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Article Title: LRRTM1 underlies synaptic convergence in visual thalamus
    Article Snippet: Gad1 1 Kb cDNA (corresponding to nucleotides 1099–2081) was generated using Superscript II Reverse Transcriptase First Strand cDNA Synthesis kit (cat # 18064014, Invitrogen, La Jolla, CA) according to the manufacturer manual, amplified by PCR using primers mentioned in the primers list, gel purified, and then cloned into a pGEM-T Easy Vector using pGEM-T Easy Vector kit, (cat # A1360, Promega, Madison, WI) according to the kit manual. .. Sense and anti-sense riboprobes against Gad1, Syt1, Nrn1 , and Lrrtm1 were synthesized from 5 µg linearized plasmids using digoxigenin-(DIG) or fluorescein-labeled uridylyltransferase (UTP) (cat # 11685619910, cat # 11277073910, Roche, Mannheim, Germany) and the MAXIscript in vitro Transcription Kit (cat # AM1312, Ambion, Austin, TX) according to the kit manual.

    Article Title: Characterization of a novel zebrafish (Danio rerio) gene, wdr81, associated with cerebellar ataxia, mental retardation and dysequilibrium syndrome (CAMRQ)
    Article Snippet: The amplicon was run on 0.8 % agarose gel, the observed band was extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), and cloned (A1360, pGEM-T Easy Vector System I, Promega, USA). .. Linearized plasmid, extracted from the gel (D4007, Zymoclean Gel DNA Recovery Kit, USA), was used as a template and antisense RNA probe was synthesized using T7 enzyme mix (AM1320, MaxiScript SP6/T7 In Vitro Transcription Kit, Ambion, USA) and DIG-labelling mix (11277073910, DIG RNA Labelling Mix, Roche, Germany).

    Transgenic Assay:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: Each mRNA cocktail was diluted in sterile buffer and injected into B6 single-cell embryos at the BUMC Transgenic Core facility ( http://www.bumc.bu.edu/transgenic/ ). .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    Produced:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: A primer set was designed (ccdBSec-Dir/ccdBSec-Rev) for sequencing plasmid inserts. pELMO digestion with the Sma I enzyme produced a blunt-ended vector; this bypassed the A-tailing reaction step for PCR fragments obtained by high fidelity polymerases. .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Genotyping Assay:

    Article Title: Hnrnph1 Is A Quantitative Trait Gene for Methamphetamine Sensitivity
    Article Snippet: We developed a genotyping assay utilizing native restriction enzyme recognition sites within the TALENs Fok I cleavage domain. .. To confirm base pair deletions in our founder lines, undigested restriction enzyme-exposed PCR amplicon bands were excised, gel-purified, and vector-ligated overnight at 4°C using the pGEM T-easy Vector Systems (Promega).

    Marker:

    Article Title: pELMO, an optimised in-house cloning vector
    Article Snippet: The pELMO vector (3306 bp) was constructed for the efficient and reliable cloning of PCR products; it contains two selection systems: the most common ampicillin-resistance marker generally used in basic research groups and the ccdB gene encoding a 101 amino acid toxic protein expressed by the lac promoter. .. Such step is essential for plasmids having T-overhangs, like pGEM-T Easy Vector Systems (Promega Corporation MD, USA).

    Gel Extraction:

    Article Title: Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
    Article Snippet: .. The amplicons were purified with an E.Z.N.A. gel extraction kit (Omega, USA) and cloned into pGEM-T Easy vector systems (Promega, USA). .. Recombinant clones were sequenced with an automatic DNA sequencer (ABI, USA) and spliced artificially.

    Article Title: Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I
    Article Snippet: .. PROTOCOLS I- Cloning of competitor Materials • RNAgents: Total RNA Isolation System, Promega Z5110 • PolyATract: mRNA Isolation System II, Promega Z5200 • Reverse Transcription System, Promega A3500 • PCR Master Mix, Promega M7505 • QIAquick Gel Extraction Kit, QIAGEN 28704 • pGEM® -T Easy Vector System I, Promega A1360 and pBluescript® II Phagemid Vectors, Stratagene 212207 to cloning DNA sequences • Reagents for cloning • Ampicillin and streptomycin for selection purposes • Degenerated primers to amplify tiGHR I probe sequence • Top 10 (F- mcrA Δ[mrr-hsdRMS-mcrBC] ø80 lacZ Δ M15 Δ lacX74deoR recA1 araD139 Δ[ara-leu] 7697 galV galK rpsL [StrR] endA1nupG) or equivalent electrocompetent E. coli cells • T7 RiboMAX TM Express RNAi System, Promega P1700 • MEGAscript® RNAi Kit, Ambion 1626 Methods 1. .. To obtain total RNA of tilapia (O. niloticus ) liver, we followed the procedure described in the section IV of the Technical Bulletin 087 (TB087) of RNAgents® Total RNA Isolation System (Promega).

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    Promega pgem t easy vector
    Pgem T Easy Vector, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 6980 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pgem t easy vector - by Bioz Stars, 2020-01
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