ecor  (New England Biolabs)


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    Name:
    EcoRV
    Description:
    EcoRV 20 000 units
    Catalog Number:
    r0195l
    Price:
    249
    Size:
    20 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs ecor
    EcoRV
    EcoRV 20 000 units
    https://www.bioz.com/result/ecor/product/New England Biolabs
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    ecor - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "Nonviral Gene Targeting at rDNA Locus of Human Mesenchymal Stem Cells"

    Article Title: Nonviral Gene Targeting at rDNA Locus of Human Mesenchymal Stem Cells

    Journal: BioMed Research International

    doi: 10.1155/2013/135189

    Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco I, EcoR I, Hind III , and Pvu II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .
    Figure Legend Snippet: Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco I, EcoR I, Hind III , and Pvu II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .

    Techniques Used: Expressing, Homologous Recombination, Produced, Polymerase Chain Reaction, Amplification, Marker, Southern Blot

    2) Product Images from "Investigation of DNA-protein Sequence-Specific Interactions with a ds-DNA Array"

    Article Title: Investigation of DNA-protein Sequence-Specific Interactions with a ds-DNA Array

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/10020417

    The images of EcoR I and Rsa I digestion on ds-DNA array. (A): the image of the ds-DNA array which created by insertion with Cy3-dUTP (a) digested by EcoR I under 1h(b) and 12h(c); (B): the image of ds-DNA array (a) digested by Rsa I under 1 h(b) and 12 h(c) digestion. The four different concentrations of Oligo II (C, 1, 2, 3 and 4 below the two images indicate control, 80 µM, 40 µM, 20 µM and 10 µM Oligo II, respectively.) were spotted onto the array.
    Figure Legend Snippet: The images of EcoR I and Rsa I digestion on ds-DNA array. (A): the image of the ds-DNA array which created by insertion with Cy3-dUTP (a) digested by EcoR I under 1h(b) and 12h(c); (B): the image of ds-DNA array (a) digested by Rsa I under 1 h(b) and 12 h(c) digestion. The four different concentrations of Oligo II (C, 1, 2, 3 and 4 below the two images indicate control, 80 µM, 40 µM, 20 µM and 10 µM Oligo II, respectively.) were spotted onto the array.

    Techniques Used: DNA Array

    The fluorescence intensity variation of EcoR I and Rsa I digestion on the same array after methylation. EM indicated methylation enzyme and ER indicated EcoR I.
    Figure Legend Snippet: The fluorescence intensity variation of EcoR I and Rsa I digestion on the same array after methylation. EM indicated methylation enzyme and ER indicated EcoR I.

    Techniques Used: Fluorescence, Methylation

    The images of array one digested by EcoR I and Rsa I after methylation. (A): Images of array one which created by inserted Cy3-dUTP (a) was treated by EcoR I methylation enzyme (b) and then by EcoR I (c); (B): Images of array one which created by inserted Cy3-dUTP (a) was treated by EcoR I methylation enzyme (b) and then by Rsa I (c). (C and E below the images indicate Control probe and Oligo I, respectively)
    Figure Legend Snippet: The images of array one digested by EcoR I and Rsa I after methylation. (A): Images of array one which created by inserted Cy3-dUTP (a) was treated by EcoR I methylation enzyme (b) and then by EcoR I (c); (B): Images of array one which created by inserted Cy3-dUTP (a) was treated by EcoR I methylation enzyme (b) and then by Rsa I (c). (C and E below the images indicate Control probe and Oligo I, respectively)

    Techniques Used: Methylation

    Related Articles

    Mutagenesis:

    Article Title: Isolation and characterization of HepP: a virulence-related Pseudomonas aeruginosa heparinase
    Article Snippet: .. Therefore, to further confirm the mutation of hepP in PA14ΔhepP , we performed restriction enzyme digestion on the PCR products with EcoR V (New England Biolabs). ..

    Isolation:

    Article Title: Regulated Expression of the Beta2-Toxin Gene (cpb2) in Clostridium perfringens Type A Isolates from Horses with Gastrointestinal Diseases
    Article Snippet: .. Isolated C. perfringens DNA samples, prepared as previously described ( , ), were digested with EcoRV or HpaI (New England Biolabs), separated by electrophoresis on 1% agarose gels, and Southern transferred. .. The blots were hybridized with the DIG-labeled cpb2 probe, which was then detected using a DIG chemiluminescence detection system utilizing CSPD [disodium 3-(4-methoxyspiro{1,2-dioxetane-3,2′-(5-chloro)tricyclo[3,3.1.13.7]decane}-4-yl)phenyl phosphate] ready-to-use substrate (Roche) as described earlier ( ).

    Purification:

    Article Title: A dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting
    Article Snippet: .. The pGRFP plasmid was cut with EcoRV (NEB), 5′ dephosphorylated with CIAP, and gel purified. .. The linearized vector was blunt-end ligated to the SU66E20 BAC library inserts using T4 DNA ligase.

    Electrophoresis:

    Article Title: Integrative and Sequence Characteristics of a Novel Genetic Element, ICE6013, in Staphylococcus aureus
    Article Snippet: .. Genomic DNA was digested with EcoRV (New England Biolabs), and restriction fragments were separated by electrophoresis in 0.5% agarose. .. The fragments were blotted onto nylon membranes (Bio-Rad) by capillary transfer.

    Article Title: Regulated Expression of the Beta2-Toxin Gene (cpb2) in Clostridium perfringens Type A Isolates from Horses with Gastrointestinal Diseases
    Article Snippet: .. Isolated C. perfringens DNA samples, prepared as previously described ( , ), were digested with EcoRV or HpaI (New England Biolabs), separated by electrophoresis on 1% agarose gels, and Southern transferred. .. The blots were hybridized with the DIG-labeled cpb2 probe, which was then detected using a DIG chemiluminescence detection system utilizing CSPD [disodium 3-(4-methoxyspiro{1,2-dioxetane-3,2′-(5-chloro)tricyclo[3,3.1.13.7]decane}-4-yl)phenyl phosphate] ready-to-use substrate (Roche) as described earlier ( ).

    Transgenic Assay:

    Article Title: Overexpression of Arabidopsis thaliana gibberellic acid 20 oxidase (AtGA20ox) gene enhance the vegetative growth and fiber quality in kenaf (Hibiscus cannabinus L.) plants
    Article Snippet: .. DNA from putative transgenic and UT plants was digested with EcoR V (NEB, UK). ..

    Molecular Weight:

    Article Title: Structural diversity of supercoiled DNA
    Article Snippet: .. BbvCI, EcoRV, Nb.BbvCI, NdeI, Nuclease Bal-31, T4 DNA Ligase, low molecular weight DNA ladder and 100 bp DNA ladder were purchased from New England Biolabs (Ipswich, MA). .. Proteinase K was purchased from Roche Molecular Biochemicals (Mannheim, Germany).

    Polymerase Chain Reaction:

    Article Title: Isolation and characterization of HepP: a virulence-related Pseudomonas aeruginosa heparinase
    Article Snippet: .. Therefore, to further confirm the mutation of hepP in PA14ΔhepP , we performed restriction enzyme digestion on the PCR products with EcoR V (New England Biolabs). ..

    Plasmid Preparation:

    Article Title: A dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting
    Article Snippet: .. The pGRFP plasmid was cut with EcoRV (NEB), 5′ dephosphorylated with CIAP, and gel purified. .. The linearized vector was blunt-end ligated to the SU66E20 BAC library inserts using T4 DNA ligase.

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    New England Biolabs ecor v
    Confirmation of the mutation in PA14Δ hepP . PA14 and PA14Δ hepP were grown in LB broth and the chromosomal DNA was extracted. a <t>PCR</t> analysis to detect the presence of MAR2xT7 within hepP . PCR reactions were run using the chromosomal DNA from each strain as a template and primers corresponding to the DNA sequences 94 bp upstream and 179 bp downstream of the hepP structural gene ( zbdP- For3/ hepP- Rev3, Table 2 ). The expected 1926-bp fragment from PA14 (lane 1) and the 2920-bp fragment (the additional 994 bp from MAR2xT7 ) from PA14Δ hepP (lane 2) were detected. Lane 3 is a no-template control and the molecular size standards are in lane 4. b Restriction analysis of the PCR products. The coding sequence for hepP does not contain an <t>EcoR</t> V restriction enzyme site, while MAR2xT7 contains a single EcoR V site. Digestion of the PCR products with EcoR V failed to reduce the size of the 1926-bp fragment obtained from PA14 (lane 3) but resulted in the cleavage of the product obtained from PA14Δ hepP into the expected 800 bp and 2120 bp fragments (lane 4). Lane 1 contains the molecular size standards; lane 2 was left empty
    Ecor V, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecor v/product/New England Biolabs
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    ecor v - by Bioz Stars, 2020-08
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    New England Biolabs ecor i
    Representative variation in MSAP profiles. “→” to red arrows represents variation in DNA methylation between diploid and tetraploid plants; “+” represents fragments obtained after digestion with <t>EcoR</t> I or Hpa II/ Msp I; “−” represents fragments not digested by EcoR I or Hpa II/ Msp I; Type I fragments are nonmethylated and were presented in both the H ( EcoR I or Hpa II digest) and M ( EcoR I or Msp I digest) lanes; Type II are fully methylated and only appeared in the M lanes; Type III are hemimethylated and appeared in the H lanes; Type IV were fragments absent from both H and M lanes in diploid but present in either H or M lane of tetraploid, and vice versa.
    Ecor I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecor i/product/New England Biolabs
    Average 99 stars, based on 55 article reviews
    Price from $9.99 to $1999.99
    ecor i - by Bioz Stars, 2020-08
    99/100 stars
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    NEBuffer Set EcoRI DpnII 5 0 ml
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    Image Search Results


    Confirmation of the mutation in PA14Δ hepP . PA14 and PA14Δ hepP were grown in LB broth and the chromosomal DNA was extracted. a PCR analysis to detect the presence of MAR2xT7 within hepP . PCR reactions were run using the chromosomal DNA from each strain as a template and primers corresponding to the DNA sequences 94 bp upstream and 179 bp downstream of the hepP structural gene ( zbdP- For3/ hepP- Rev3, Table 2 ). The expected 1926-bp fragment from PA14 (lane 1) and the 2920-bp fragment (the additional 994 bp from MAR2xT7 ) from PA14Δ hepP (lane 2) were detected. Lane 3 is a no-template control and the molecular size standards are in lane 4. b Restriction analysis of the PCR products. The coding sequence for hepP does not contain an EcoR V restriction enzyme site, while MAR2xT7 contains a single EcoR V site. Digestion of the PCR products with EcoR V failed to reduce the size of the 1926-bp fragment obtained from PA14 (lane 3) but resulted in the cleavage of the product obtained from PA14Δ hepP into the expected 800 bp and 2120 bp fragments (lane 4). Lane 1 contains the molecular size standards; lane 2 was left empty

    Journal: BMC Microbiology

    Article Title: Isolation and characterization of HepP: a virulence-related Pseudomonas aeruginosa heparinase

    doi: 10.1186/s12866-017-1141-0

    Figure Lengend Snippet: Confirmation of the mutation in PA14Δ hepP . PA14 and PA14Δ hepP were grown in LB broth and the chromosomal DNA was extracted. a PCR analysis to detect the presence of MAR2xT7 within hepP . PCR reactions were run using the chromosomal DNA from each strain as a template and primers corresponding to the DNA sequences 94 bp upstream and 179 bp downstream of the hepP structural gene ( zbdP- For3/ hepP- Rev3, Table 2 ). The expected 1926-bp fragment from PA14 (lane 1) and the 2920-bp fragment (the additional 994 bp from MAR2xT7 ) from PA14Δ hepP (lane 2) were detected. Lane 3 is a no-template control and the molecular size standards are in lane 4. b Restriction analysis of the PCR products. The coding sequence for hepP does not contain an EcoR V restriction enzyme site, while MAR2xT7 contains a single EcoR V site. Digestion of the PCR products with EcoR V failed to reduce the size of the 1926-bp fragment obtained from PA14 (lane 3) but resulted in the cleavage of the product obtained from PA14Δ hepP into the expected 800 bp and 2120 bp fragments (lane 4). Lane 1 contains the molecular size standards; lane 2 was left empty

    Article Snippet: Therefore, to further confirm the mutation of hepP in PA14ΔhepP , we performed restriction enzyme digestion on the PCR products with EcoR V (New England Biolabs).

    Techniques: Mutagenesis, Polymerase Chain Reaction, Sequencing

    Identification of the recombinant plasmid. (A) Detection of the housekeeping gene GAPDH. (B) Identification of the target gene by RT-PCR. (C) PCR product of the Fyn gene was subcloned into pMD18-T-Fyn and pEGFP-N1-Fyn. (D) Identification of pMD18-T-Fyn fragments produced by restriction enzyme digestion with Eco R I and Sma I. (E) Identification of pEGFP-N1-Fyn fragments generated by restriction enzyme digestion with Eco R I and Sma I. Lane 1, RT-PCR GAPDH product; Lane 2, negative control; Lane 3, RT-PCR Fyn product from brain; Lane 4, positive control; Lane 5, PCR product of pMD18-T-Fyn; Lane 6, PCR product of pEGFP-N1-Fyn; Lanes 7 and 9, positive control; Lanes 8 and 10, recombinant plasmid identification by digestion with EcoR I and Sma I (pMD18-T-Fyn, 2700 bp; pEGFP-N1, 4700 bp; Fyn, 1611 bp); Lane M, DNA marker.

    Journal: Journal of Veterinary Science

    Article Title: Mouse Fyn induces pseudopodium formation in Chinese hamster ovary cells

    doi: 10.4142/jvs.2014.15.1.111

    Figure Lengend Snippet: Identification of the recombinant plasmid. (A) Detection of the housekeeping gene GAPDH. (B) Identification of the target gene by RT-PCR. (C) PCR product of the Fyn gene was subcloned into pMD18-T-Fyn and pEGFP-N1-Fyn. (D) Identification of pMD18-T-Fyn fragments produced by restriction enzyme digestion with Eco R I and Sma I. (E) Identification of pEGFP-N1-Fyn fragments generated by restriction enzyme digestion with Eco R I and Sma I. Lane 1, RT-PCR GAPDH product; Lane 2, negative control; Lane 3, RT-PCR Fyn product from brain; Lane 4, positive control; Lane 5, PCR product of pMD18-T-Fyn; Lane 6, PCR product of pEGFP-N1-Fyn; Lanes 7 and 9, positive control; Lanes 8 and 10, recombinant plasmid identification by digestion with EcoR I and Sma I (pMD18-T-Fyn, 2700 bp; pEGFP-N1, 4700 bp; Fyn, 1611 bp); Lane M, DNA marker.

    Article Snippet: Restriction enzymes EcoR I and Sma I were purchased from New England Biolabs (USA).

    Techniques: Recombinant, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Produced, Generated, Negative Control, Positive Control, Marker

    Selection of putative CaMV35S : AtGA20ox transgenic kenaf plants in MS media containing hygromycin B and the confirmation through PCR amplification and Southern blot analysis. Growth of UT plants in the MS medium without hygromycin B (a) and with hygromycin B (45 μg mL −1 ) (b). Growing of putative transformants in the MS media with hygromycin B (45 μg mL −1 ) (c and d). Detection of the transgene encoding AtGA20ox under duplicate 35S promoter in transgenic G4 and V36 kenaf and UT plants either amplified by PCR using GA-R and 35S-F primers (e and f) or southern blot analysis (g and h). Lane M is ladder, lanes 1–3, 5, 7 (e) and 1–3, 5, 13 (f) are putative transformed G4 and V36 plants respectively showing two amplifications 1.3 kb and 1.6 kb, except for line G4-1 (lane 1 in e) which produced only 1.3 kb amplification product; lane UT is the DNA of untransformed plants; lane −Ve is negative control used PCR product without DNA; lane +Ve is the plasmid DNA of expression clone pEXP32-AtGA20ox. Southern blot analysis was performed with genomic DNA (6 μg–30 μg) digested with EcoR V, separated on 0.8% agarose gel using biotin labelled ORF of AtGA20ox as probe: lane M is marker; lanes 1–3, 5, 7 are putative G4 transformants; lane UT is the DNA of G4 UT and lane +Ve is plasmid DNA (g). In Figure h, lanes 1, 3, 13 are the putative V36 transformants, lane UT is the DNA from V36 UT plant and lane +Ve is plasmid DNA of pEXP32-AtGA20ox digested with EcoR V.

    Journal: Breeding Science

    Article Title: Overexpression of Arabidopsis thaliana gibberellic acid 20 oxidase (AtGA20ox) gene enhance the vegetative growth and fiber quality in kenaf (Hibiscus cannabinus L.) plants

    doi: 10.1270/jsbbs.65.177

    Figure Lengend Snippet: Selection of putative CaMV35S : AtGA20ox transgenic kenaf plants in MS media containing hygromycin B and the confirmation through PCR amplification and Southern blot analysis. Growth of UT plants in the MS medium without hygromycin B (a) and with hygromycin B (45 μg mL −1 ) (b). Growing of putative transformants in the MS media with hygromycin B (45 μg mL −1 ) (c and d). Detection of the transgene encoding AtGA20ox under duplicate 35S promoter in transgenic G4 and V36 kenaf and UT plants either amplified by PCR using GA-R and 35S-F primers (e and f) or southern blot analysis (g and h). Lane M is ladder, lanes 1–3, 5, 7 (e) and 1–3, 5, 13 (f) are putative transformed G4 and V36 plants respectively showing two amplifications 1.3 kb and 1.6 kb, except for line G4-1 (lane 1 in e) which produced only 1.3 kb amplification product; lane UT is the DNA of untransformed plants; lane −Ve is negative control used PCR product without DNA; lane +Ve is the plasmid DNA of expression clone pEXP32-AtGA20ox. Southern blot analysis was performed with genomic DNA (6 μg–30 μg) digested with EcoR V, separated on 0.8% agarose gel using biotin labelled ORF of AtGA20ox as probe: lane M is marker; lanes 1–3, 5, 7 are putative G4 transformants; lane UT is the DNA of G4 UT and lane +Ve is plasmid DNA (g). In Figure h, lanes 1, 3, 13 are the putative V36 transformants, lane UT is the DNA from V36 UT plant and lane +Ve is plasmid DNA of pEXP32-AtGA20ox digested with EcoR V.

    Article Snippet: DNA from putative transgenic and UT plants was digested with EcoR V (NEB, UK).

    Techniques: Selection, Transgenic Assay, Mass Spectrometry, Polymerase Chain Reaction, Amplification, Southern Blot, Transformation Assay, Produced, Negative Control, Plasmid Preparation, Expressing, Agarose Gel Electrophoresis, Marker

    Representative variation in MSAP profiles. “→” to red arrows represents variation in DNA methylation between diploid and tetraploid plants; “+” represents fragments obtained after digestion with EcoR I or Hpa II/ Msp I; “−” represents fragments not digested by EcoR I or Hpa II/ Msp I; Type I fragments are nonmethylated and were presented in both the H ( EcoR I or Hpa II digest) and M ( EcoR I or Msp I digest) lanes; Type II are fully methylated and only appeared in the M lanes; Type III are hemimethylated and appeared in the H lanes; Type IV were fragments absent from both H and M lanes in diploid but present in either H or M lane of tetraploid, and vice versa.

    Journal: International Journal of Molecular Sciences

    Article Title: Morphological, Genome and Gene Expression Changes in Newly Induced Autopolyploid Chrysanthemum lavandulifolium (Fisch. ex Trautv.) Makino

    doi: 10.3390/ijms17101690

    Figure Lengend Snippet: Representative variation in MSAP profiles. “→” to red arrows represents variation in DNA methylation between diploid and tetraploid plants; “+” represents fragments obtained after digestion with EcoR I or Hpa II/ Msp I; “−” represents fragments not digested by EcoR I or Hpa II/ Msp I; Type I fragments are nonmethylated and were presented in both the H ( EcoR I or Hpa II digest) and M ( EcoR I or Msp I digest) lanes; Type II are fully methylated and only appeared in the M lanes; Type III are hemimethylated and appeared in the H lanes; Type IV were fragments absent from both H and M lanes in diploid but present in either H or M lane of tetraploid, and vice versa.

    Article Snippet: Mthylation Sensitive Amplified Polymorphism Analysis The MSAP technique was applied to the DNA pools from three diploid lines and three tetraploid lines C. lavandulifolium They were digested with either EcoR I and Hpa II or EcoR I and Msp I (NEB) at 37 °C for 12 h. The digested fragments were ligated to 5 pmol EcoR I adaptor and 50 pmol Hpa II/Msp I adaptor by incubation with 4 U T4 DNA polymerase (NEB) at 16 °C for 4 has described for the AFLP method.

    Techniques: DNA Methylation Assay, Methylation