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Stratagene competent e coli xl1 blue cells
Competent E Coli Xl1 Blue Cells, supplied by Stratagene, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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competent e coli xl1 blue cells - by Bioz Stars, 2020-04
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Acetylene Reduction Assay:

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: PCR for amplification of rbcL was performed with primer pairs rbcL 1 ([ ] and rbcL R, 5′-CCA WCG CAT ARA NGG TTG HGA-3′ ([ ] modifid after [ ]) by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 53°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 45°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. tufA amplification was performed with primer pair tufAF and tufAR [ ] by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 57°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 48°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. PCR products were size-fractionated in a 1.5% agarose gel for 90 min at 70 V and bands according to desired gene-fragment length and subsequently gel-extracted using Machery-Nagel NucleoSpin® Extract II (Düren, Germany) kit following manufacturer’s instructions. .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

Amplification:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: The CYP51 gene of C. albicans DUMC136 was amplified as two partially overlapping fragments. .. The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: Paragraph title: Rapid amplification of cDNA 3′ ends. ... The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies).

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: Isolation of Blo t 2 isoforms Full length Blo t 2 was amplified from B. tropicalis cDNA using primers Blo t 2 LIC F: 5′-GACGACGACAAGATCATGTTCAAGTTTATCTGTCTC-3′ and Blot 2 LIC R: 5′-GAGGAGAAGCCCGGTTTAATCGACAACCTCGG-3′ with highly thermostable pfu polymerase. .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: Paragraph title: DNA amplification ... Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: .. The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. Positive (white-colony morphotype) transformants were streaked for isolation and stored in both stab cultures and dimethyl sulfoxide.

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Selected DNA fragments were amplified by PCR and then cloned into the pGEM-T Easy vector (Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The amplified product was isolated, restricted, and ligated into the expression vector pT7-7 ( ). .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Paragraph title: Amplification of phage particles ... Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method.

Clone Assay:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: Paragraph title: Cloning and nucleotide sequencing of CYP51. ... The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies). .. Plasmids from 22 E. coli colonies derived from wild-type and 24 colonies derived from Spt5 ΔCTR clones, respectively, were sequenced (GATC Biotech) using pET28b sequencing primers (forward, 5′-TAATACGACTCACTATAGGG-3′; reverse, 5′-GGGTTATGCTAGTTATTGC-3′).

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: The cloning of mIDO-1 (GenBank accession number ) and mIDO-2 (GenBank accession number ) was described previously ( , ). .. Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed.

Article Title: Nitric Oxide Inhibits Rhinovirus-Induced Cytokine Production and Viral Replication in a Human Respiratory Epithelial Cell Line
Article Snippet: .. The full-length cDNA was cloned into a pCR II vector (Invitrogen Corp., San Diego, Calif.) between two Eco RI sites and grown in competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. The sequence of the cDNA probe used for Northern analysis was identical to the published sequence of IL-8 ( , ), as determined by dideoxy sequencing.

Article Title: In Vivo Analysis of Human T-Cell Leukemia Virus Type 1 Reverse Transcription Accuracy
Article Snippet: Purified genomic DNA ( ) from pools of step 3 clones was digested with the restriction enzyme Sal I to release the neo , pACYC origin of replication, and lacZ α peptide gene sequences from the HTLV-1 shuttle vector proviral DNA (Fig. ). .. The purified proviral DNA was ligated and used to electroporate competent E. coli XL1 Blue cells (Stratagene).

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany). .. For each specimen 12 clones were sequenced by Macrogen Inc, Amsterdam.

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. Clones were numbered discontinuously from 1 to 182 and assigned the prefix OCS (for Oregon coast study).

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Selected DNA fragments were amplified by PCR and then cloned into the pGEM-T Easy vector (Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Construct:

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. Phage stocks of the different Avd display constructs were made from individual colonies picked from the transformation plates into super broth (SB) medium supplemented with the appropriate antibiotics and glucose.

Incubation:

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: The enzyme was inactivated by incubation at 65°C for 20 min. PCRs were conducted using the cDNA samples, 0.25 μM gene-specific upstream primer containing an EcoRI restriction site (underlined in the sequence 5′-TAT GAATTC TTCTGTTTTGGGTTTGGA-3′), an anchored primer (5′-GA CTCGAG TCGACATCGA-3′; an XhoI restriction site is underlined), and Taq DNA polymerase (Fermentas; Thermo Fisher Scientific). .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies).

Expressing:

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The amplified product was isolated, restricted, and ligated into the expression vector pT7-7 ( ). .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: Paragraph title: 2.1. Expression and Purification of Recombinant Amelogenins and rpMmp-20 ... The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA).

Western Blot:

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. Phages were PEG precipitated and analyzed by SDS-PAGE and western blotting following immunostaining with the polyclonal rabbit α-avd IgG (University of Oulu, 1:5000) and the monoclonal mouse anti-pIII IgG (Biosite, Sweden, 1:2000) antibodies.

Transformation Assay:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: .. The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene). .. Sequencing was carried out with an ABI Prism Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin-Elmer).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies). .. Plasmids from 22 E. coli colonies derived from wild-type and 24 colonies derived from Spt5 ΔCTR clones, respectively, were sequenced (GATC Biotech) using pET28b sequencing primers (forward, 5′-TAATACGACTCACTATAGGG-3′; reverse, 5′-GGGTTATGCTAGTTATTGC-3′).

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. One hundred and forty colonies were picked, inoculated overnight and plasmid extracted.

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: .. Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed. .. After the correct mutations were confirmed by sequencing, the genes were cloned into pDEST17.

Article Title: The Flp double cross system a simple efficient procedure for cloning DNA fragments
Article Snippet: Competent E. coli XL1-Blue cells (recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacI q Z ΔM15 Tn10 (Tetr )]) were purchased from Stratagene (La Jolla, CA, USA, cat. # 200249). .. Transformation and staining colonies for beta galactosidase with X-Gal were done according to the manufacturer's instructions.

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM). .. Positive clones were randomly selected using white/blue screening and were sequenced on an automated ABI 3500xL Genetic Analyzer (Applied Biosystems, Foster City, California, USA) using T7 and SP6 primers and a BigDye Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems).

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: .. The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA). .. Recombinant pig enamelysin (rpMmp-20) was expressed in 500ml media from the cells and purified following the protocol described previously ( , ).

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: .. Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. Phage stocks of the different Avd display constructs were made from individual colonies picked from the transformation plates into super broth (SB) medium supplemented with the appropriate antibiotics and glucose.

Derivative Assay:

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies). .. Plasmids from 22 E. coli colonies derived from wild-type and 24 colonies derived from Spt5 ΔCTR clones, respectively, were sequenced (GATC Biotech) using pET28b sequencing primers (forward, 5′-TAATACGACTCACTATAGGG-3′; reverse, 5′-GGGTTATGCTAGTTATTGC-3′).

Hybridization:

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: An enrichment was performed using hybridization-based capture with (GT)8 and (CT)8 biotinylated probes and streptavidin-coated magnetic beads (Streptavidin Magnesphere Paramagnetic Particles, Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

High Performance Liquid Chromatography:

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: Briefly, rP172 and rP148 were purified by ammonium sulfate precipitation and reverse phase high performance liquid chromatography (RP-HPLC), using buffer A (0.1% trifluoroacetic acid: TFA) and buffer B (buffer A +60% acetonitrile). .. The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA).

Electroporation:

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM). .. Positive clones were randomly selected using white/blue screening and were sequenced on an automated ABI 3500xL Genetic Analyzer (Applied Biosystems, Foster City, California, USA) using T7 and SP6 primers and a BigDye Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems).

Chromatography:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: Immunoglobulin G (IgG) obtained by protein A chromatography (Hi-Trap protein A; Amersham-Pharmacia Biotech) was applied to the TcTS affinity column, and after several washings with saline, specific antibodies were acid eluted. .. After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures.

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene). .. KDO8P synthase from N. gonorrhoeae was overexpressed in E. coli BL21(DE3) cells harboring pT7-7-NGKDO and purified, as previously reported, using anion-exchange chromatography ( ; W. P. Taylor and R. W. Woodard, submitted for publication).

Ligation:

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: .. The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. Positive (white-colony morphotype) transformants were streaked for isolation and stored in both stab cultures and dimethyl sulfoxide.

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene). .. Plasmids isolated from several transformants were subjected to DNA sequencing, and one plasmid with the expected sequence was isolated (pT7-7-NGKDO).

Library Screening:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: Paragraph title: Peptide combinatorial library screening. ... After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures.

Genomic Sequencing:

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM). .. All obtained sequences were analyzed to identify microsatellite-enriched regions with the Simple Sequence Repeat Identification Tool (SSRIT) [ ], and oligonucleotides complementary to genomic sequences flanking the microsatellite region were designed using Primer3Plus [ ] with the following criteria: preferable primer size between 18 and 22 bp; melting temperature (Tm) between 50 and 60 °C; amplified product length between 100 and 300 bp; and GC content between 40 and 60%.

Synthesized:

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: The ACT1 cDNA was synthesized as described previously ( ) using SuperScript II reverse transcriptase (Invitrogen), 0.5 μM anchored oligo(dT) primer containing an XhoI restriction site, underlined in the sequence 5′-GA CTCGAG TCGACATCGATTTTTTTTTTTTTTTTT-3′, and 2 μg of RNA template. .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies).

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM). .. Following these criteria, 56 primer pairs were designed and synthesized for analysis.

Northern Blot:

Article Title: Nitric Oxide Inhibits Rhinovirus-Induced Cytokine Production and Viral Replication in a Human Respiratory Epithelial Cell Line
Article Snippet: Paragraph title: Probes for Northern blotting. ... The full-length cDNA was cloned into a pCR II vector (Invitrogen Corp., San Diego, Calif.) between two Eco RI sites and grown in competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.).

Infection:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: Sera from T. cruzi (CAI and RA strains)-infected rabbits were taken 3 months after infection. .. After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures.

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. The bacterial cultures were infected with the helper phage (1012 pfu/ml) VCS-M13 (Stratagene, LaJolla, CA) for amplification of Avd phages.

Generated:

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: On the basis of a detailed analysis of the heme environment in mIDO-2 in comparison to the crystal structure of hIDO-1 (described below), three mutants of mIDO-2 were generated via site-directed mutagenesis: mIDO-2 with an M-to-L change at position 385 (mIDO-2M385L ), mIDO-2 with an Y-to-F change at position 231 (mIDO-2Y231F ), and mIDO-2 with an A-to-D change at position 384 (mIDO-2A384D ). .. Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed.

DNA Sequencing:

Article Title: In Vivo Analysis of Human T-Cell Leukemia Virus Type 1 Reverse Transcription Accuracy
Article Snippet: Paragraph title: Recovery of shuttle vector proviral DNA and DNA sequencing of the lacZ α peptide region. ... The purified proviral DNA was ligated and used to electroporate competent E. coli XL1 Blue cells (Stratagene).

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene). .. Plasmids isolated from several transformants were subjected to DNA sequencing, and one plasmid with the expected sequence was isolated (pT7-7-NGKDO).

Methylation:

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: .. Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed. .. After the correct mutations were confirmed by sequencing, the genes were cloned into pDEST17.

Sequencing:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: Paragraph title: Cloning and nucleotide sequencing of CYP51. ... The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: The enzyme was inactivated by incubation at 65°C for 20 min. PCRs were conducted using the cDNA samples, 0.25 μM gene-specific upstream primer containing an EcoRI restriction site (underlined in the sequence 5′-TAT GAATTC TTCTGTTTTGGGTTTGGA-3′), an anchored primer (5′-GA CTCGAG TCGACATCGA-3′; an XhoI restriction site is underlined), and Taq DNA polymerase (Fermentas; Thermo Fisher Scientific). .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies).

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. The plasmid DNA was then submitted to double pass sequencing, and all sequences were analysed using DNAMAN® (Lynnon Corp., Quebec, Canada).

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed. .. After the correct mutations were confirmed by sequencing, the genes were cloned into pDEST17.

Article Title: Nitric Oxide Inhibits Rhinovirus-Induced Cytokine Production and Viral Replication in a Human Respiratory Epithelial Cell Line
Article Snippet: The full-length cDNA was cloned into a pCR II vector (Invitrogen Corp., San Diego, Calif.) between two Eco RI sites and grown in competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. The sequence of the cDNA probe used for Northern analysis was identical to the published sequence of IL-8 ( , ), as determined by dideoxy sequencing.

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Paragraph title: Construction of microsatellite-enriched libraries and sequence analysis ... Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: Standard PCR methodologies were employed to amplify this gene sequence from N. gonorrhoeae DNA. .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Affinity Purification:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures. .. Filters were screened with TcTS affinity-purified immunoglobulins from T. cruzi -infected rabbits.

Binding Assay:

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA). .. A His-TRAP chelating nickel affinity column (Pharmacia, Piscataway, NJ,USA) with a three buffer system (Loading buffer A: 6M Guanidine-HCl, 0.01M Tris-HCl, 0.1M NaH2 PO4 , pH=8.0, binding buffer B: 8M urea, 0.1M NaH2 PO4 and 0.01 M Tris-HCl, pH=8.0 and eluting buffer C: 8M urea, 0.1 M NaH2PO4, 0.01M Tris-HCl, 0.25M imidazole, pH= 6.3) was used to isolate the recombinant protein from E-coli products.

Magnetic Beads:

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: An enrichment was performed using hybridization-based capture with (GT)8 and (CT)8 biotinylated probes and streptavidin-coated magnetic beads (Streptavidin Magnesphere Paramagnetic Particles, Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Mutagenesis:

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: Paragraph title: Site-directed mutagenesis of mIDO-2. ... Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed.

Article Title: In Vivo Analysis of Human T-Cell Leukemia Virus Type 1 Reverse Transcription Accuracy
Article Snippet: The purified proviral DNA was ligated and used to electroporate competent E. coli XL1 Blue cells (Stratagene). .. The ratio of white plus light-blue bacterial colonies to total bacterial colonies observed provided a forward mutant frequency for a single retroviral replication cycle.

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: Full-length recombinant pig amelogenin (rP172) and an engineered mutant form lacking the hydrophilic C-terminal 24 amino acids (rP148) were expressed in e-coli and purified as previously described ( , ). .. The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA).

Isolation:

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: Paragraph title: Isolation of Blo t 2 isoforms ... The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany). .. For each specimen 12 clones were sequenced by Macrogen Inc, Amsterdam.

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. Positive (white-colony morphotype) transformants were streaked for isolation and stored in both stab cultures and dimethyl sulfoxide.

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The amplified product was isolated, restricted, and ligated into the expression vector pT7-7 ( ). .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Purification:

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: Supernatants from four washings with 100 μl of PBS were pooled, and 5 μg of purified antibodies was added and allowed to react for 2 h in a final volume of 550 μl. .. After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures.

Article Title: In Vivo Analysis of Human T-Cell Leukemia Virus Type 1 Reverse Transcription Accuracy
Article Snippet: .. The purified proviral DNA was ligated and used to electroporate competent E. coli XL1 Blue cells (Stratagene). ..

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene). .. KDO8P synthase from N. gonorrhoeae was overexpressed in E. coli BL21(DE3) cells harboring pT7-7-NGKDO and purified, as previously reported, using anion-exchange chromatography ( ; W. P. Taylor and R. W. Woodard, submitted for publication).

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: Paragraph title: 2.1. Expression and Purification of Recombinant Amelogenins and rpMmp-20 ... The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA).

Polymerase Chain Reaction:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: .. The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene). .. Sequencing was carried out with an ABI Prism Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin-Elmer).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies). .. Plasmids from 22 E. coli colonies derived from wild-type and 24 colonies derived from Spt5 ΔCTR clones, respectively, were sequenced (GATC Biotech) using pET28b sequencing primers (forward, 5′-TAATACGACTCACTATAGGG-3′; reverse, 5′-GGGTTATGCTAGTTATTGC-3′).

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Benzo[b]quinolizinium Derivatives Have a Strong Antimalarial Activity and Inhibit Indoleamine Dioxygenase
Article Snippet: Mutations were introduced into mIDO-2 by PCR with Pfu DNA polymerase (Promega, USA) using mutated primers (for mIDO-2Y231F , sense primer 5′-CCCAGACATATTTT T CTCGGTCATCC-3′ and antisense primer 5′-GGATGACCGAG A AAAATATGTCTGGG-3′; for mIDO-2A384D , sense primer 5′-GGGTACTG A CATGCTGAGCTTCTTG-3′ and antisense primer 5′-CAAGAAGCTCAGCATG T CAGTACCC-3′; for mIDO-2M385L , sense primer 5′-GTACTGCC C TGCTGAGCTTCTTGAA-3′ and antisense primer 5′-TTCAAGAAGCTCAGCA G GGCAGTAC-3′; the mutated nucleotide is underlined). .. Methylated nonmutated template plasmids were digested with DpnI, and competent Escherichia coli XL1-Blue cells (Stratagene) were subsequently transformed.

Article Title: Nitric Oxide Inhibits Rhinovirus-Induced Cytokine Production and Viral Replication in a Human Respiratory Epithelial Cell Line
Article Snippet: .. The full-length cDNA was cloned into a pCR II vector (Invitrogen Corp., San Diego, Calif.) between two Eco RI sites and grown in competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. The sequence of the cDNA probe used for Northern analysis was identical to the published sequence of IL-8 ( , ), as determined by dideoxy sequencing.

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: PCR for amplification of rbcL was performed with primer pairs rbcL 1 ([ ] and rbcL R, 5′-CCA WCG CAT ARA NGG TTG HGA-3′ ([ ] modifid after [ ]) by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 53°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 45°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. tufA amplification was performed with primer pair tufAF and tufAR [ ] by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 57°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 48°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. PCR products were size-fractionated in a 1.5% agarose gel for 90 min at 70 V and bands according to desired gene-fragment length and subsequently gel-extracted using Machery-Nagel NucleoSpin® Extract II (Düren, Germany) kit following manufacturer’s instructions. .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: SSU rRNA genes were amplified from the environmental sample of genomic DNAs by PCR ( ) with two general bacterial SSU rDNA primers and Pfu DNA polymerase as previously described ( ). .. The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.).

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Selected DNA fragments were amplified by PCR and then cloned into the pGEM-T Easy vector (Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: Standard PCR methodologies were employed to amplify this gene sequence from N. gonorrhoeae DNA. .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Positron Emission Tomography:

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. One hundred and forty colonies were picked, inoculated overnight and plasmid extracted.

Immunostaining:

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. Phages were PEG precipitated and analyzed by SDS-PAGE and western blotting following immunostaining with the polyclonal rabbit α-avd IgG (University of Oulu, 1:5000) and the monoclonal mouse anti-pIII IgG (Biosite, Sweden, 1:2000) antibodies.

Chloramphenicol Acetyltransferase Assay:

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: PCR for amplification of rbcL was performed with primer pairs rbcL 1 ([ ] and rbcL R, 5′-CCA WCG CAT ARA NGG TTG HGA-3′ ([ ] modifid after [ ]) by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 53°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 45°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. tufA amplification was performed with primer pair tufAF and tufAR [ ] by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 57°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 48°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. PCR products were size-fractionated in a 1.5% agarose gel for 90 min at 70 V and bands according to desired gene-fragment length and subsequently gel-extracted using Machery-Nagel NucleoSpin® Extract II (Düren, Germany) kit following manufacturer’s instructions. .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

SDS Page:

Article Title: Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein
Article Snippet: Phagemid vectors with the Avd insert were transformed into chemically competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA) with the heat shock method. .. Phages were PEG precipitated and analyzed by SDS-PAGE and western blotting following immunostaining with the polyclonal rabbit α-avd IgG (University of Oulu, 1:5000) and the monoclonal mouse anti-pIII IgG (Biosite, Sweden, 1:2000) antibodies.

Plasmid Preparation:

Article Title: Formation of Azole-Resistant Candida albicans by Mutation of Sterol 14-Demethylase P450
Article Snippet: .. The PCR products were ligated into Sma I-cut pBluescript II SK(+) vector (Stratagene) and transformed into competent Escherichia coli XL1-Blue cells (Stratagene). .. Sequencing was carried out with an ABI Prism Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin-Elmer).

Article Title: The Spt5 C-Terminal Region Recruits Yeast 3? RNA Cleavage Factor I
Article Snippet: .. The PCR product and the pET28b vector were digested with EcoRI-HF as well as XhoI (New England BioLabs, Inc.) and then ligated and transformed into competent Escherichia coli XL1-Blue cells (Stratagene; Agilent Technologies). .. Plasmids from 22 E. coli colonies derived from wild-type and 24 colonies derived from Spt5 ΔCTR clones, respectively, were sequenced (GATC Biotech) using pET28b sequencing primers (forward, 5′-TAATACGACTCACTATAGGG-3′; reverse, 5′-GGGTTATGCTAGTTATTGC-3′).

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Nitric Oxide Inhibits Rhinovirus-Induced Cytokine Production and Viral Replication in a Human Respiratory Epithelial Cell Line
Article Snippet: .. The full-length cDNA was cloned into a pCR II vector (Invitrogen Corp., San Diego, Calif.) between two Eco RI sites and grown in competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. The sequence of the cDNA probe used for Northern analysis was identical to the published sequence of IL-8 ( , ), as determined by dideoxy sequencing.

Article Title: In Vivo Analysis of Human T-Cell Leukemia Virus Type 1 Reverse Transcription Accuracy
Article Snippet: Paragraph title: Recovery of shuttle vector proviral DNA and DNA sequencing of the lacZ α peptide region. ... The purified proviral DNA was ligated and used to electroporate competent E. coli XL1 Blue cells (Stratagene).

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany). .. For each specimen 12 clones were sequenced by Macrogen Inc, Amsterdam.

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: .. The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.). .. Positive (white-colony morphotype) transformants were streaked for isolation and stored in both stab cultures and dimethyl sulfoxide.

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: Selected DNA fragments were amplified by PCR and then cloned into the pGEM-T Easy vector (Promega, Madison, Wisconsin, USA). .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM).

Article Title: Substrate Ambiguity of 3-Deoxy-d-manno-Octulosonate 8-Phosphate Synthase from Neisseria gonorrhoeae Revisited
Article Snippet: The amplified product was isolated, restricted, and ligated into the expression vector pT7-7 ( ). .. The ligation mixture was used to transform competent E. coli XL1-Blue cells (Stratagene).

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: .. The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA). .. Recombinant pig enamelysin (rpMmp-20) was expressed in 500ml media from the cells and purified following the protocol described previously ( , ).

Multiplex Assay:

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: 2.5 μl of genomic DNA was used as template in a 20 μl final volume reaction supplied with 5.5 μl sterilized water, 2 μl Qiagen® Q-Solution, 10 μl of double concentrated QIAGEN® Multiplex PCR Master Mix and 1 μl of 5 pmol/μl concentrated primer each. .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

Recombinant:

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. One hundred and forty colonies were picked, inoculated overnight and plasmid extracted.

Article Title: First microsatellite markers for Paspalum plicatulum (Poaceae) characterization and cross-amplification in different Paspalum species of the Plicatula group
Article Snippet: .. Competent Escherichia coli XL1-Blue cells (Stratagene, Agilent Technologies, Santa Clara, California, USA) were transformed with recombinant plasmids via the electroporation method and were then cultivated on agar medium containing ampicillin (100 mg/ml), X-galactosidase 2% (100 µg/ml) and IPTG (100 mM). .. Positive clones were randomly selected using white/blue screening and were sequenced on an automated ABI 3500xL Genetic Analyzer (Applied Biosystems, Foster City, California, USA) using T7 and SP6 primers and a BigDye Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems).

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: .. The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA). .. Recombinant pig enamelysin (rpMmp-20) was expressed in 500ml media from the cells and purified following the protocol described previously ( , ).

Agarose Gel Electrophoresis:

Article Title: Blo t 2: Group 2 allergen from the dust mite Blomia tropicalis
Article Snippet: The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene). .. The recombinant pET-32(a) vectors were transformed into competent E. coli XL1-Blue cells (Stratagene).

Article Title: Identification of sequestered chloroplasts in photosynthetic and non-photosynthetic sacoglossan sea slugs (Mollusca, Gastropoda)
Article Snippet: PCR for amplification of rbcL was performed with primer pairs rbcL 1 ([ ] and rbcL R, 5′-CCA WCG CAT ARA NGG TTG HGA-3′ ([ ] modifid after [ ]) by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 53°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 45°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. tufA amplification was performed with primer pair tufAF and tufAR [ ] by an initially denaturation for 15 min at 95°C, followed by 9 touch-down cycles at 94°C for 45 s, 57°C (-1°C per cycle) for 45 s, 72°C for 90 s, followed by 25 standard cycles (94°C for 45 s, 48°C for 45 s and 72°C for 90 s.) and a final extension at 72°C for 10 min. PCR products were size-fractionated in a 1.5% agarose gel for 90 min at 70 V and bands according to desired gene-fragment length and subsequently gel-extracted using Machery-Nagel NucleoSpin® Extract II (Düren, Germany) kit following manufacturer’s instructions. .. Isolated fragments were ligated into pGEM T-easy Vector (Promega, Germany) and cloned into competent E. coli XL1-blue cells from Stratagene (Heidelberg, Germany).

Concentration Assay:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: .. After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures. .. Filters were screened with TcTS affinity-purified immunoglobulins from T. cruzi -infected rabbits.

Staining:

Article Title: The Flp double cross system a simple efficient procedure for cloning DNA fragments
Article Snippet: Competent E. coli XL1-Blue cells (recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacI q Z ΔM15 Tn10 (Tetr )]) were purchased from Stratagene (La Jolla, CA, USA, cat. # 200249). .. Transformation and staining colonies for beta galactosidase with X-Gal were done according to the manufacturer's instructions.

Environmental Sampling:

Article Title: Phylogenetic Diversity of Ultraplankton Plastid Small-Subunit rRNA Genes Recovered in Environmental Nucleic Acid Samples from the Pacific and Atlantic Coasts of the United States
Article Snippet: SSU rRNA genes were amplified from the environmental sample of genomic DNAs by PCR ( ) with two general bacterial SSU rDNA primers and Pfu DNA polymerase as previously described ( ). .. The amplification products from six reactions were pooled, inserted into the Sma I restriction site of phagemid vector pBluescript KSII− (Stratagene) by blunt-end ligation as previously described ( , ), and used to transform competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.).

Affinity Column:

Article Title: Epitope Mapping of trans-Sialidase from Trypanosoma cruzi Reveals the Presence of Several Cross-Reactive Determinants
Article Snippet: Immunoglobulin G (IgG) obtained by protein A chromatography (Hi-Trap protein A; Amersham-Pharmacia Biotech) was applied to the TcTS affinity column, and after several washings with saline, specific antibodies were acid eluted. .. After 10 washings with 1 ml of PBS and one more with 0.15 M NaCl, beads were resuspended in 500 μl of glycine–HCl (concentration, 1 M; pH 2.5) for 30 min. Tris base (50 μl of a 1 M solution) was added, and the suspension was immediately used to infect 1 ml of competent Escherichia coli XL1-Blue cells (Stratagene, La Jolla, Calif.); then the techniques described in detail by Felici et al. ( ) were carefully followed in the phage rescue and filter immunoscreening procedures.

Article Title: Amelogenin "Nanorods" Formation During Proteolysis by Mmp-20
Article Snippet: The recombinant enamelysin plasmid (provided by Dr. James P. Simmer) was transformed into competent E. coli XL1-Blue cells (Stratagene, La Jolla, CA, USA). .. A His-TRAP chelating nickel affinity column (Pharmacia, Piscataway, NJ,USA) with a three buffer system (Loading buffer A: 6M Guanidine-HCl, 0.01M Tris-HCl, 0.1M NaH2 PO4 , pH=8.0, binding buffer B: 8M urea, 0.1M NaH2 PO4 and 0.01 M Tris-HCl, pH=8.0 and eluting buffer C: 8M urea, 0.1 M NaH2PO4, 0.01M Tris-HCl, 0.25M imidazole, pH= 6.3) was used to isolate the recombinant protein from E-coli products.

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    Stratagene e coli xl1 blue competent cells
    E Coli Xl1 Blue Competent Cells, supplied by Stratagene, used in various techniques. Bioz Stars score: 87/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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