ecor1 new england biolabs  (New England Biolabs)


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    Name:
    NEBuffer EcoR I
    Description:
    NEBuffer EcoR I 6 0 ml
    Catalog Number:
    b0101s
    Price:
    20
    Size:
    6 0 ml
    Category:
    DNA Manipulation
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    Structured Review

    New England Biolabs ecor1 new england biolabs
    NEBuffer EcoR I
    NEBuffer EcoR I 6 0 ml
    https://www.bioz.com/result/ecor1 new england biolabs/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ecor1 new england biolabs - by Bioz Stars, 2020-08
    94/100 stars

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    Related Articles

    Amplification:

    Article Title: ILDR2: An Endoplasmic Reticulum Resident Molecule Mediating Hepatic Lipid Homeostasis
    Article Snippet: .. The destination vector and the PCR amplified Ildr2 sequence were digested with HindIII and EcoR1 (NEBiolabs) in NEBuffer EcoR1 and BSA at 37°C for 60 min, purified and ligated. .. C-terminal -tagged ILDR2 mYFP construct.

    Isolation:

    Article Title: Autoimmune Regulator (AIRE) Is Expressed in Spermatogenic Cells, and It Altered the Expression of Several Nucleic-Acid-Binding and Cytoskeletal Proteins in Germ Cell 1 Spermatogonial (GC1-spg) Cells *
    Article Snippet: .. Lipofectamine 2000, DMEM, DMEMF12, OptiMEM, FBS, antibiotic–antimycotic mixture l -glutamine, 1X MEM vitamins, nonessential amino acids StemPro34 SFM base (Invitrogen, Green Island, CA); EcoR1, Streptomyces albus 1 (Sal1), calf intestinal phosphatase (CIP), New England Biolab (NEB) buffer 3, EcoR1 Buffer, DNA ligase, and ligase buffer (New England Biolabs, Ipswich, MA); Nucleobond Xtra midi Plus EF kit (Macherey Nagel, Duren, Germany); gel extraction kit (GE Healthcare, Buckinghamshire, UK); 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), MgCl2 , KCl, NaCl, 2,2′,2″,2‴-(ethane-1,2-diyldinitrilo) tetra acetic acid (EDTA), glycerol, SUPERSCRIPT® VILOTM cDNA synthesis kit (Invitrogen, Waltham, MA); SYBR green master mix plates, ABI PRISM® 384-well optical reaction (Applied Bio systems, Waltham, MA), QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany); protease inhibitor mixture, bovine serum albumin (Calbiochem, San Diego, CA); Total RNA isolation (TRI) reagent, glucose, d-biotin, insulin, pyruvic acid sodium salt, dl-lactic acid, ascorbic acid, sodium selenite, putrescine, bovine Apo-transferrin, progesterone, β-estradiol 17-cypionate, 2-mercaptoethanol (Sigma- Aldrich, Carlsbad, MO); fetal bovine serum (Hyclone, Logan, UT); SCP-3, histone H3 (Abcam, Cambridge, United Kingdom); anti-Aire antibody (sc-33188), AIRE-1 (M-300), GFRα-1 (H-70), OCT-3/4 (H-134), protamine 2 (C-14), PGK2 (F-25), actin (I-19), goat anti-rabbit HRP, donkey anti-goat HRP, and HRP-conjugated anti-rabbit secondary antibody (sc-2317, Santa Cruz Biotechnology, Dallas, TX) were procured. .. Mus musculus, Swiss albino strain, housed and inbred at the Laboratory Animal Research Centre (LARC) of Rajiv Gandhi Centre for Biotechnology, Trivandrum, India, were used.

    Purification:

    Article Title: Mechanisms underlying the impact of humic acids on DNA quantification by SYBR Green I and consequences for the analysis of soils and aquatic sediments
    Article Snippet: .. The plasmid was purified by applying the Qiaprep Spin Miniprep kit from Qiagen (Hilden, Germany). pACYC 184 was linearised by Eco RI restriction endonuclease digestion in NEBuffer Eco RI (New England Biolabs) according to the supplier’s protocol and purified by applying the QIAquick PCR purification kit from Qiagen. ..

    Article Title: ILDR2: An Endoplasmic Reticulum Resident Molecule Mediating Hepatic Lipid Homeostasis
    Article Snippet: .. The destination vector and the PCR amplified Ildr2 sequence were digested with HindIII and EcoR1 (NEBiolabs) in NEBuffer EcoR1 and BSA at 37°C for 60 min, purified and ligated. .. C-terminal -tagged ILDR2 mYFP construct.

    SYBR Green Assay:

    Article Title: Autoimmune Regulator (AIRE) Is Expressed in Spermatogenic Cells, and It Altered the Expression of Several Nucleic-Acid-Binding and Cytoskeletal Proteins in Germ Cell 1 Spermatogonial (GC1-spg) Cells *
    Article Snippet: .. Lipofectamine 2000, DMEM, DMEMF12, OptiMEM, FBS, antibiotic–antimycotic mixture l -glutamine, 1X MEM vitamins, nonessential amino acids StemPro34 SFM base (Invitrogen, Green Island, CA); EcoR1, Streptomyces albus 1 (Sal1), calf intestinal phosphatase (CIP), New England Biolab (NEB) buffer 3, EcoR1 Buffer, DNA ligase, and ligase buffer (New England Biolabs, Ipswich, MA); Nucleobond Xtra midi Plus EF kit (Macherey Nagel, Duren, Germany); gel extraction kit (GE Healthcare, Buckinghamshire, UK); 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), MgCl2 , KCl, NaCl, 2,2′,2″,2‴-(ethane-1,2-diyldinitrilo) tetra acetic acid (EDTA), glycerol, SUPERSCRIPT® VILOTM cDNA synthesis kit (Invitrogen, Waltham, MA); SYBR green master mix plates, ABI PRISM® 384-well optical reaction (Applied Bio systems, Waltham, MA), QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany); protease inhibitor mixture, bovine serum albumin (Calbiochem, San Diego, CA); Total RNA isolation (TRI) reagent, glucose, d-biotin, insulin, pyruvic acid sodium salt, dl-lactic acid, ascorbic acid, sodium selenite, putrescine, bovine Apo-transferrin, progesterone, β-estradiol 17-cypionate, 2-mercaptoethanol (Sigma- Aldrich, Carlsbad, MO); fetal bovine serum (Hyclone, Logan, UT); SCP-3, histone H3 (Abcam, Cambridge, United Kingdom); anti-Aire antibody (sc-33188), AIRE-1 (M-300), GFRα-1 (H-70), OCT-3/4 (H-134), protamine 2 (C-14), PGK2 (F-25), actin (I-19), goat anti-rabbit HRP, donkey anti-goat HRP, and HRP-conjugated anti-rabbit secondary antibody (sc-2317, Santa Cruz Biotechnology, Dallas, TX) were procured. .. Mus musculus, Swiss albino strain, housed and inbred at the Laboratory Animal Research Centre (LARC) of Rajiv Gandhi Centre for Biotechnology, Trivandrum, India, were used.

    Polymerase Chain Reaction:

    Article Title: Mechanisms underlying the impact of humic acids on DNA quantification by SYBR Green I and consequences for the analysis of soils and aquatic sediments
    Article Snippet: .. The plasmid was purified by applying the Qiaprep Spin Miniprep kit from Qiagen (Hilden, Germany). pACYC 184 was linearised by Eco RI restriction endonuclease digestion in NEBuffer Eco RI (New England Biolabs) according to the supplier’s protocol and purified by applying the QIAquick PCR purification kit from Qiagen. ..

    Article Title: ILDR2: An Endoplasmic Reticulum Resident Molecule Mediating Hepatic Lipid Homeostasis
    Article Snippet: .. The destination vector and the PCR amplified Ildr2 sequence were digested with HindIII and EcoR1 (NEBiolabs) in NEBuffer EcoR1 and BSA at 37°C for 60 min, purified and ligated. .. C-terminal -tagged ILDR2 mYFP construct.

    Incubation:

    Article Title: The teneurin C-terminal domain possesses nuclease activity and is apoptogenic
    Article Snippet: .. We incubated 200 ng of pcDNA3 (Thermo Fisher Scientific) for 2 h with various concentrations of the teneurin-1 CTD in the presence of 1× EcoRI buffer (New England BioLabs, Ipswich, USA, B0101S) to determine the optimal conditions for further studies ( D). .. Further reactions included the pcDNA3 plasmid mixed with 500 ng of the CTDs of chicken teneurins-1 and -2 at 37°C for different periods of time, predigestion of the plasmid with EcoRI (stopping the reaction by heating to 65°C for 20 min prior to the addition of the purified teneurin domains), and including 10 mM EDTA in the reaction mix.

    Article Title: The teneurin C-terminal domain possesses nuclease activity and is apoptogenic
    Article Snippet: .. Endonuclease activity We incubated 200 ng of pcDNA3 (Thermo Fisher Scientific) for 2 h with various concentrations of the teneurin-1 CTD in the presence of 1× EcoRI buffer (New England BioLabs, Ipswich, USA, B0101S) to determine the optimal conditions for further studies ( D). .. Further reactions included the pcDNA3 plasmid mixed with 500 ng of the CTDs of chicken teneurins-1 and -2 at 37°C for different periods of time, predigestion of the plasmid with EcoRI (stopping the reaction by heating to 65°C for 20 min prior to the addition of the purified teneurin domains), and including 10 mM EDTA in the reaction mix.

    Activity Assay:

    Article Title: The teneurin C-terminal domain possesses nuclease activity and is apoptogenic
    Article Snippet: .. Endonuclease activity We incubated 200 ng of pcDNA3 (Thermo Fisher Scientific) for 2 h with various concentrations of the teneurin-1 CTD in the presence of 1× EcoRI buffer (New England BioLabs, Ipswich, USA, B0101S) to determine the optimal conditions for further studies ( D). .. Further reactions included the pcDNA3 plasmid mixed with 500 ng of the CTDs of chicken teneurins-1 and -2 at 37°C for different periods of time, predigestion of the plasmid with EcoRI (stopping the reaction by heating to 65°C for 20 min prior to the addition of the purified teneurin domains), and including 10 mM EDTA in the reaction mix.

    Protease Inhibitor:

    Article Title: Autoimmune Regulator (AIRE) Is Expressed in Spermatogenic Cells, and It Altered the Expression of Several Nucleic-Acid-Binding and Cytoskeletal Proteins in Germ Cell 1 Spermatogonial (GC1-spg) Cells *
    Article Snippet: .. Lipofectamine 2000, DMEM, DMEMF12, OptiMEM, FBS, antibiotic–antimycotic mixture l -glutamine, 1X MEM vitamins, nonessential amino acids StemPro34 SFM base (Invitrogen, Green Island, CA); EcoR1, Streptomyces albus 1 (Sal1), calf intestinal phosphatase (CIP), New England Biolab (NEB) buffer 3, EcoR1 Buffer, DNA ligase, and ligase buffer (New England Biolabs, Ipswich, MA); Nucleobond Xtra midi Plus EF kit (Macherey Nagel, Duren, Germany); gel extraction kit (GE Healthcare, Buckinghamshire, UK); 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), MgCl2 , KCl, NaCl, 2,2′,2″,2‴-(ethane-1,2-diyldinitrilo) tetra acetic acid (EDTA), glycerol, SUPERSCRIPT® VILOTM cDNA synthesis kit (Invitrogen, Waltham, MA); SYBR green master mix plates, ABI PRISM® 384-well optical reaction (Applied Bio systems, Waltham, MA), QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany); protease inhibitor mixture, bovine serum albumin (Calbiochem, San Diego, CA); Total RNA isolation (TRI) reagent, glucose, d-biotin, insulin, pyruvic acid sodium salt, dl-lactic acid, ascorbic acid, sodium selenite, putrescine, bovine Apo-transferrin, progesterone, β-estradiol 17-cypionate, 2-mercaptoethanol (Sigma- Aldrich, Carlsbad, MO); fetal bovine serum (Hyclone, Logan, UT); SCP-3, histone H3 (Abcam, Cambridge, United Kingdom); anti-Aire antibody (sc-33188), AIRE-1 (M-300), GFRα-1 (H-70), OCT-3/4 (H-134), protamine 2 (C-14), PGK2 (F-25), actin (I-19), goat anti-rabbit HRP, donkey anti-goat HRP, and HRP-conjugated anti-rabbit secondary antibody (sc-2317, Santa Cruz Biotechnology, Dallas, TX) were procured. .. Mus musculus, Swiss albino strain, housed and inbred at the Laboratory Animal Research Centre (LARC) of Rajiv Gandhi Centre for Biotechnology, Trivandrum, India, were used.

    Sequencing:

    Article Title: ILDR2: An Endoplasmic Reticulum Resident Molecule Mediating Hepatic Lipid Homeostasis
    Article Snippet: .. The destination vector and the PCR amplified Ildr2 sequence were digested with HindIII and EcoR1 (NEBiolabs) in NEBuffer EcoR1 and BSA at 37°C for 60 min, purified and ligated. .. C-terminal -tagged ILDR2 mYFP construct.

    Gel Extraction:

    Article Title: Autoimmune Regulator (AIRE) Is Expressed in Spermatogenic Cells, and It Altered the Expression of Several Nucleic-Acid-Binding and Cytoskeletal Proteins in Germ Cell 1 Spermatogonial (GC1-spg) Cells *
    Article Snippet: .. Lipofectamine 2000, DMEM, DMEMF12, OptiMEM, FBS, antibiotic–antimycotic mixture l -glutamine, 1X MEM vitamins, nonessential amino acids StemPro34 SFM base (Invitrogen, Green Island, CA); EcoR1, Streptomyces albus 1 (Sal1), calf intestinal phosphatase (CIP), New England Biolab (NEB) buffer 3, EcoR1 Buffer, DNA ligase, and ligase buffer (New England Biolabs, Ipswich, MA); Nucleobond Xtra midi Plus EF kit (Macherey Nagel, Duren, Germany); gel extraction kit (GE Healthcare, Buckinghamshire, UK); 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), MgCl2 , KCl, NaCl, 2,2′,2″,2‴-(ethane-1,2-diyldinitrilo) tetra acetic acid (EDTA), glycerol, SUPERSCRIPT® VILOTM cDNA synthesis kit (Invitrogen, Waltham, MA); SYBR green master mix plates, ABI PRISM® 384-well optical reaction (Applied Bio systems, Waltham, MA), QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany); protease inhibitor mixture, bovine serum albumin (Calbiochem, San Diego, CA); Total RNA isolation (TRI) reagent, glucose, d-biotin, insulin, pyruvic acid sodium salt, dl-lactic acid, ascorbic acid, sodium selenite, putrescine, bovine Apo-transferrin, progesterone, β-estradiol 17-cypionate, 2-mercaptoethanol (Sigma- Aldrich, Carlsbad, MO); fetal bovine serum (Hyclone, Logan, UT); SCP-3, histone H3 (Abcam, Cambridge, United Kingdom); anti-Aire antibody (sc-33188), AIRE-1 (M-300), GFRα-1 (H-70), OCT-3/4 (H-134), protamine 2 (C-14), PGK2 (F-25), actin (I-19), goat anti-rabbit HRP, donkey anti-goat HRP, and HRP-conjugated anti-rabbit secondary antibody (sc-2317, Santa Cruz Biotechnology, Dallas, TX) were procured. .. Mus musculus, Swiss albino strain, housed and inbred at the Laboratory Animal Research Centre (LARC) of Rajiv Gandhi Centre for Biotechnology, Trivandrum, India, were used.

    Plasmid Preparation:

    Article Title: Mechanisms underlying the impact of humic acids on DNA quantification by SYBR Green I and consequences for the analysis of soils and aquatic sediments
    Article Snippet: .. The plasmid was purified by applying the Qiaprep Spin Miniprep kit from Qiagen (Hilden, Germany). pACYC 184 was linearised by Eco RI restriction endonuclease digestion in NEBuffer Eco RI (New England Biolabs) according to the supplier’s protocol and purified by applying the QIAquick PCR purification kit from Qiagen. ..

    Article Title: ILDR2: An Endoplasmic Reticulum Resident Molecule Mediating Hepatic Lipid Homeostasis
    Article Snippet: .. The destination vector and the PCR amplified Ildr2 sequence were digested with HindIII and EcoR1 (NEBiolabs) in NEBuffer EcoR1 and BSA at 37°C for 60 min, purified and ligated. .. C-terminal -tagged ILDR2 mYFP construct.

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  • 94
    New England Biolabs ecor1
    (A) Colony PCR analysis of yeast clones obtained from H1 and H4 genomic DNA by the method shown in Figure 1B . Ligated samples (see legend, Figure 2 ) were used to transform S.cerevisiae . In contrast to results with E.coli ( Figure 2 ), full-length candidate clones were obtained. Lanes with dots are from colonies used in further analyses (see text). The DNA ladder is as in Figures 1 and 2 . ( B ) Verification of clone structure. To confirm the structure of the full-length candidates, yeast minipreps were treated with <t>EcoR1</t> and subjected to random-primed rolling circle amplification with phi-29 DNA polymerase. As an example, H4#4 is displayed on a 2.5% agarose gel after diagnostic digestion with Xbal and PvuII. Upper and lower arrows indicate the vector backbone and insert bands, respectively. ( C ) Efficacy of the EcoR1 pre-digestion. The EcoR1 pre-treated sample in B (‘+’) is run on a 1% agarose gel alongside an untreated (‘−’) miniprep of H4#4. Treated and untreated samples were phi-29 amplified in parallel and digested with XbaI and PvuII. Bands evident in the untreated lane correspond in size to those predicted for 2 µ circle DNA. Bands observed with EcoR1 pre-treatment (arrows) correspond to the pYes2.1 vector and insert.
    Ecor1, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecor1/product/New England Biolabs
    Average 94 stars, based on 71 article reviews
    Price from $9.99 to $1999.99
    ecor1 - by Bioz Stars, 2020-08
    94/100 stars
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    99
    New England Biolabs ecori
    Final genotypes in a binary format for subset of the samples and alleles from the AFLP run with the primer pair <t>FAM-Ecori-ACA</t> and <t>Msei-CTT</t> as displayed in the Genotypes tab of the GeneMapper software.
    Ecori, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 985 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecori/product/New England Biolabs
    Average 99 stars, based on 985 article reviews
    Price from $9.99 to $1999.99
    ecori - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    (A) Colony PCR analysis of yeast clones obtained from H1 and H4 genomic DNA by the method shown in Figure 1B . Ligated samples (see legend, Figure 2 ) were used to transform S.cerevisiae . In contrast to results with E.coli ( Figure 2 ), full-length candidate clones were obtained. Lanes with dots are from colonies used in further analyses (see text). The DNA ladder is as in Figures 1 and 2 . ( B ) Verification of clone structure. To confirm the structure of the full-length candidates, yeast minipreps were treated with EcoR1 and subjected to random-primed rolling circle amplification with phi-29 DNA polymerase. As an example, H4#4 is displayed on a 2.5% agarose gel after diagnostic digestion with Xbal and PvuII. Upper and lower arrows indicate the vector backbone and insert bands, respectively. ( C ) Efficacy of the EcoR1 pre-digestion. The EcoR1 pre-treated sample in B (‘+’) is run on a 1% agarose gel alongside an untreated (‘−’) miniprep of H4#4. Treated and untreated samples were phi-29 amplified in parallel and digested with XbaI and PvuII. Bands evident in the untreated lane correspond in size to those predicted for 2 µ circle DNA. Bands observed with EcoR1 pre-treatment (arrows) correspond to the pYes2.1 vector and insert.

    Journal: Nucleic Acids Research

    Article Title: New approaches to the analysis of palindromic sequences from the human genome: evolution and polymorphism of an intronic site at the NF1 locus

    doi: 10.1093/nar/gni189

    Figure Lengend Snippet: (A) Colony PCR analysis of yeast clones obtained from H1 and H4 genomic DNA by the method shown in Figure 1B . Ligated samples (see legend, Figure 2 ) were used to transform S.cerevisiae . In contrast to results with E.coli ( Figure 2 ), full-length candidate clones were obtained. Lanes with dots are from colonies used in further analyses (see text). The DNA ladder is as in Figures 1 and 2 . ( B ) Verification of clone structure. To confirm the structure of the full-length candidates, yeast minipreps were treated with EcoR1 and subjected to random-primed rolling circle amplification with phi-29 DNA polymerase. As an example, H4#4 is displayed on a 2.5% agarose gel after diagnostic digestion with Xbal and PvuII. Upper and lower arrows indicate the vector backbone and insert bands, respectively. ( C ) Efficacy of the EcoR1 pre-digestion. The EcoR1 pre-treated sample in B (‘+’) is run on a 1% agarose gel alongside an untreated (‘−’) miniprep of H4#4. Treated and untreated samples were phi-29 amplified in parallel and digested with XbaI and PvuII. Bands evident in the untreated lane correspond in size to those predicted for 2 µ circle DNA. Bands observed with EcoR1 pre-treatment (arrows) correspond to the pYes2.1 vector and insert.

    Article Snippet: EcoR1 gave differential cleavage (i.e. cut the 2 micron circle and spared the plasmid clone) eliminating almost all amplification of the competing 2 micron circle DNA ( ).

    Techniques: Polymerase Chain Reaction, Clone Assay, Random Primed, Amplification, Agarose Gel Electrophoresis, Diagnostic Assay, Plasmid Preparation

    ( A ) Map (to scale) of the palindromic region within a 3.8 kb EcoR1 fragment of the NF1 gene. Exons are numbered according to L05367 (see text). ‘R1’ denotes EcoR1 sites according to Southern blot data ( 27 ) and the reference human genome sequence (May 2004). ( B ) Cloning strategy. Oligos used for PCR flank the palindromic site. The PCR product is ligated into a commercial vector (see Materials and Methods). ‘X’ and ‘P’ refer to the Xbal and PvuII sites used in diagnostic digests. ( C ) PCR amplification of various DNA templates. Lane ‘M’, markers (Trackit 100 bp ladder, Invitrogen). ‘B’ is a PCR with Bac clone CTD-2370N5; ‘H1’ to ‘H5’ are with human genomic DNA from the indicated individuals. ‘C’ is with chimpanzee DNA. ‘φH4’ and ‘φG’ are from an aliquot of H4 and gorilla genomic DNA that had first been amplified with phi-29 polymerase. The H4 samples demonstrate reproducibility of the PCR as well as the fidelity of phi-29 amplification.

    Journal: Nucleic Acids Research

    Article Title: New approaches to the analysis of palindromic sequences from the human genome: evolution and polymorphism of an intronic site at the NF1 locus

    doi: 10.1093/nar/gni189

    Figure Lengend Snippet: ( A ) Map (to scale) of the palindromic region within a 3.8 kb EcoR1 fragment of the NF1 gene. Exons are numbered according to L05367 (see text). ‘R1’ denotes EcoR1 sites according to Southern blot data ( 27 ) and the reference human genome sequence (May 2004). ( B ) Cloning strategy. Oligos used for PCR flank the palindromic site. The PCR product is ligated into a commercial vector (see Materials and Methods). ‘X’ and ‘P’ refer to the Xbal and PvuII sites used in diagnostic digests. ( C ) PCR amplification of various DNA templates. Lane ‘M’, markers (Trackit 100 bp ladder, Invitrogen). ‘B’ is a PCR with Bac clone CTD-2370N5; ‘H1’ to ‘H5’ are with human genomic DNA from the indicated individuals. ‘C’ is with chimpanzee DNA. ‘φH4’ and ‘φG’ are from an aliquot of H4 and gorilla genomic DNA that had first been amplified with phi-29 polymerase. The H4 samples demonstrate reproducibility of the PCR as well as the fidelity of phi-29 amplification.

    Article Snippet: EcoR1 gave differential cleavage (i.e. cut the 2 micron circle and spared the plasmid clone) eliminating almost all amplification of the competing 2 micron circle DNA ( ).

    Techniques: Southern Blot, Sequencing, Clone Assay, Polymerase Chain Reaction, Plasmid Preparation, Diagnostic Assay, Amplification, BAC Assay

    Figure 1. Expression of bioactive chemokines from plasmid encoded mucosal chemokine and the induction of infiltration, after intramuscular injection, by cognate receptor positive cells ( A ). All genes were cloned into the multiple cloning region of the expression vector pVAX (kanamycin resistance) using the restriction enzyme sites, EcoR1 and Xho1. ( B ) Expression of pCCL25 was confirmed by a T7 coupled transcription/translation reticulocyte lysate system. The blotting gel shows size markers (designated M, lane 1), pVAX background control protein (V, lane 2), 14.2 kDa CCL25 protein (lane 3). ( C ) Expression of bioactive chemokine protein translated from the plasmid forms of CCL25. ELISA was performed using supernatants from pCCL25-transfected RD cells (pg/ml). Vector background control is included (gray bar) vs. pCCL25 (black bar). Data are shown as pg/ml of chemokine protein ± SD of triplicate wells. ( D ) Infiltration of CCR9 positive cells induced following intramuscular injection of 100 μg of pCCL25. Immunohistochemical staining of quadriceps sections 7 d post immunization with 100 μg of vector backbone control (left panel) or pCCL25. Infiltrating cells were enumerated following a visual count by microscope of brown positive cells over 6 (20×) fields, and the total number of CCR9+ cells was averaged and shown as ± SD for vector (gray bar) and pCCL25 injection (black bar). ( E ) Frequency of CD11c+ cells that express CCR9 in the popliteal and inguinal DLNs of individual immunized mice in vector backbone immunized (solid black symbols) vs. pCCL25 immunized mice (open circles). (*p

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Generation of antigen-specific immunity following systemic immunization with DNA vaccine encoding CCL25 chemokine immunoadjuvant

    doi: 10.4161/hv.22574

    Figure Lengend Snippet: Figure 1. Expression of bioactive chemokines from plasmid encoded mucosal chemokine and the induction of infiltration, after intramuscular injection, by cognate receptor positive cells ( A ). All genes were cloned into the multiple cloning region of the expression vector pVAX (kanamycin resistance) using the restriction enzyme sites, EcoR1 and Xho1. ( B ) Expression of pCCL25 was confirmed by a T7 coupled transcription/translation reticulocyte lysate system. The blotting gel shows size markers (designated M, lane 1), pVAX background control protein (V, lane 2), 14.2 kDa CCL25 protein (lane 3). ( C ) Expression of bioactive chemokine protein translated from the plasmid forms of CCL25. ELISA was performed using supernatants from pCCL25-transfected RD cells (pg/ml). Vector background control is included (gray bar) vs. pCCL25 (black bar). Data are shown as pg/ml of chemokine protein ± SD of triplicate wells. ( D ) Infiltration of CCR9 positive cells induced following intramuscular injection of 100 μg of pCCL25. Immunohistochemical staining of quadriceps sections 7 d post immunization with 100 μg of vector backbone control (left panel) or pCCL25. Infiltrating cells were enumerated following a visual count by microscope of brown positive cells over 6 (20×) fields, and the total number of CCR9+ cells was averaged and shown as ± SD for vector (gray bar) and pCCL25 injection (black bar). ( E ) Frequency of CD11c+ cells that express CCR9 in the popliteal and inguinal DLNs of individual immunized mice in vector backbone immunized (solid black symbols) vs. pCCL25 immunized mice (open circles). (*p

    Article Snippet: The 435 (CCL25) base pair PCR products were respectively ligated into pVAX1 cloning vector following a restriction enzyme digestion using EcoR1 and Xho1 (New England Biolabs) restriction sites.

    Techniques: Expressing, Plasmid Preparation, Injection, Clone Assay, Enzyme-linked Immunosorbent Assay, Transfection, Immunohistochemistry, Staining, Microscopy, Mouse Assay

    Final genotypes in a binary format for subset of the samples and alleles from the AFLP run with the primer pair FAM-Ecori-ACA and Msei-CTT as displayed in the Genotypes tab of the GeneMapper software.

    Journal: Journal of Biomolecular Techniques : JBT

    Article Title: Identification of Hedysarum Varieties Using Amplified Fragment Length Polymorphism on a Capillary Electrophoresis System

    doi:

    Figure Lengend Snippet: Final genotypes in a binary format for subset of the samples and alleles from the AFLP run with the primer pair FAM-Ecori-ACA and Msei-CTT as displayed in the Genotypes tab of the GeneMapper software.

    Article Snippet: Briefly, genomic DNA was digested with the restriction enzymes EcoR1 (New England Biolabs #R0101S) and MseI (New England Biolabs #R0525S), ligated to adapters using T4 DNA Ligase (New England Biolabs #M0202S), and used in a preselective amplification step using the AFLP Ligation/Preselective Amplification Module (Applied Biosystems P/N 402004).

    Techniques: Software