pbr322 dna msp  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Name:
    pBR322 DNA Msp I Digest
    Description:
    pBR322 DNA Msp I Digest 250 gel lanes
    Catalog Number:
    n3032l
    Price:
    302
    Size:
    250 gel lanes
    Category:
    DNA Molecular Weight Markers
    Buy from Supplier


    Structured Review

    New England Biolabs pbr322 dna msp
    pBR322 DNA Msp I Digest
    pBR322 DNA Msp I Digest 250 gel lanes
    https://www.bioz.com/result/pbr322 dna msp/product/New England Biolabs
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    pbr322 dna msp - by Bioz Stars, 2020-05
    93/100 stars

    Images

    1) Product Images from "Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor ?1 subunit"

    Article Title: Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor ?1 subunit

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2006.106617

    Cells with fast initial decay kinetics express β 1 subunit mRNA A and B , scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B , represents a recording dominated by brief and large IPSCs ( ). C , ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A 1 value than A 2 whereas the opposite is true for the slower cell. D , ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β 1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the Msp I digest of pBR322. The size of each fragment is noted on the right. The parameters for β 1 negative cells in lane 4: A 1 = 9.80, τ 1 = 13.75, A 2 = 14.17, τ 2 = 64.61, τ DW = 43.82l; and lane 5: A 1 = 12.94, τ 1 = 12.94, A 2 = 5.69, τ 2 = 77.21, τ DW = 29.63. The β 1 positive cell in lane 6: A 1 = 18.05, τ 1 = 15.11, A 2 = 4.15, τ 2 = 74.44, τ DW = 26.20.
    Figure Legend Snippet: Cells with fast initial decay kinetics express β 1 subunit mRNA A and B , scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B , represents a recording dominated by brief and large IPSCs ( ). C , ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A 1 value than A 2 whereas the opposite is true for the slower cell. D , ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β 1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the Msp I digest of pBR322. The size of each fragment is noted on the right. The parameters for β 1 negative cells in lane 4: A 1 = 9.80, τ 1 = 13.75, A 2 = 14.17, τ 2 = 64.61, τ DW = 43.82l; and lane 5: A 1 = 12.94, τ 1 = 12.94, A 2 = 5.69, τ 2 = 77.21, τ DW = 29.63. The β 1 positive cell in lane 6: A 1 = 18.05, τ 1 = 15.11, A 2 = 4.15, τ 2 = 74.44, τ DW = 26.20.

    Techniques Used: Isolation, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control, Polymerase Chain Reaction, Molecular Weight, Marker

    2) Product Images from "Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization"

    Article Title: Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    Pre-gel hybridization parameters. ( A ) Schematic diagram of pre-gel hybridization technique. ( B ) Comparison of 1 pmol each of biotinylated PNA and DNA pBR322 15-mer probes hybridized to 2 ng of Bst NI-digested pBR322. Lanes: 1, PNA probe; 2, DNA probe. ( C ) The effect of different sodium ion concentrations on a mixture of Msp I (50 ng) and Bst NI (15 ng) digests of pBR322. One picomole of fluorescently labeled PNA was used. Lanes: 1, no salt; 2, 0.1 mM; 3, 0.5 mM; 4, 1 mM; 5, 5 mM; 6, 10 mM; 7, 20 mM; 8, 30 mM; 9, 40 mM; 10, 50 mM; 11, 75 mM; 12, 100 mM; 13, 250 mM; and 14, 500 mM. ( D ) Size separation hybridizing 1 pmol lambda fluorescein-tagged PNA probe and 50 ng of each digest. Lanes: 1, Hin dIII/ Bam HI; 2, Eco RI/ Bam HI; 3, Bst EII; 4, Hin dIII.
    Figure Legend Snippet: Pre-gel hybridization parameters. ( A ) Schematic diagram of pre-gel hybridization technique. ( B ) Comparison of 1 pmol each of biotinylated PNA and DNA pBR322 15-mer probes hybridized to 2 ng of Bst NI-digested pBR322. Lanes: 1, PNA probe; 2, DNA probe. ( C ) The effect of different sodium ion concentrations on a mixture of Msp I (50 ng) and Bst NI (15 ng) digests of pBR322. One picomole of fluorescently labeled PNA was used. Lanes: 1, no salt; 2, 0.1 mM; 3, 0.5 mM; 4, 1 mM; 5, 5 mM; 6, 10 mM; 7, 20 mM; 8, 30 mM; 9, 40 mM; 10, 50 mM; 11, 75 mM; 12, 100 mM; 13, 250 mM; and 14, 500 mM. ( D ) Size separation hybridizing 1 pmol lambda fluorescein-tagged PNA probe and 50 ng of each digest. Lanes: 1, Hin dIII/ Bam HI; 2, Eco RI/ Bam HI; 3, Bst EII; 4, Hin dIII.

    Techniques Used: Hybridization, Labeling

    CE separation of a mixed Msp I- and Bst NI-digested pBR322. ( A ) UV detection. Voltage: 5 kV. Detection: 260 nm. Buffer: TBE/7 M urea/0.5% PEO (8 MDa). ( B ) LIF detection. Voltage: 5 kV. Detection: LIF 488 nm/520 nm. Buffer: 1× TBE/1% PEO (4 MDa). Temperature 50°C.
    Figure Legend Snippet: CE separation of a mixed Msp I- and Bst NI-digested pBR322. ( A ) UV detection. Voltage: 5 kV. Detection: 260 nm. Buffer: TBE/7 M urea/0.5% PEO (8 MDa). ( B ) LIF detection. Voltage: 5 kV. Detection: LIF 488 nm/520 nm. Buffer: 1× TBE/1% PEO (4 MDa). Temperature 50°C.

    Techniques Used: Multiple Displacement Amplification

    3) Product Images from "ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile"

    Article Title: ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    Journal: Molecular Biology International

    doi: 10.1155/2012/283974

    ASGR1 transcript profiles in different individuals. Nested PCR was used to detect ASGR1 transcripts T1 and T2 (first round primers A1CDSF1 + A1CDSR1; second round A1TEST2 + ASG1RTR). A product of 408 bp represented T1, present in RNA from the M + L cell fraction of five individuals (A–E) and also in HepG2 and THP1; a 291 bp product indicated the presence of T2 (individual E, and, very faintly, in HepG2). The RT-PCR product for individual A was coloaded with the DNA size standard (pBR322/ Msp I, 238 bp and 120 bp sizes indicated). Analysis was done using agarose gel electrophoresis. The figure is a composite of various data (indicated by individual gel windows).
    Figure Legend Snippet: ASGR1 transcript profiles in different individuals. Nested PCR was used to detect ASGR1 transcripts T1 and T2 (first round primers A1CDSF1 + A1CDSR1; second round A1TEST2 + ASG1RTR). A product of 408 bp represented T1, present in RNA from the M + L cell fraction of five individuals (A–E) and also in HepG2 and THP1; a 291 bp product indicated the presence of T2 (individual E, and, very faintly, in HepG2). The RT-PCR product for individual A was coloaded with the DNA size standard (pBR322/ Msp I, 238 bp and 120 bp sizes indicated). Analysis was done using agarose gel electrophoresis. The figure is a composite of various data (indicated by individual gel windows).

    Techniques Used: Nested PCR, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis

    ASGR2 transcript profiles in different individuals. Primer pair ASGR2RTF3 + ASGR2RTR2 gave RT-PCR products of different length (arrowed) according to the ASGR2 transcripts present. RNA extracted from the M + L cell fraction of 6 different individuals (A–F) gave either a profile consistent with transcripts encoding all isoforms (a, b, c, and d) or just with isoforms b and d. The liver cell line HepG2 showed all transcripts, the monocyte cell line THP1 showed only transcripts encoding isoforms b and d. The analysis was done using polyacrylamide gel electrophoresis. DNA size reference was pBR322/ Msp I (“Marker”, 238 bp and 120 bp sizes indicated). In addition to the expected products, additional major bands were visualized on polyacrylamide gel electrophoresis of the RT-PCR screen for ASGR2 transcripts (bracketed bands). It was considered possible that these represented heteroduplexes arising by cross-hybridization of complementary strands of the closely homologous true PCR products. To test this, each candidate heteroduplex band was excised from the gel, eluted into water, reamplified using the primers used in the initial PCR and the products electrophoresed on polyacrylamide. The results confirmed that the additional bands were hybrid duplexes containing complementary strands from nonidentical true PCR products: each hybrid yielded products consistent with amplification of two different template strands (data not shown).
    Figure Legend Snippet: ASGR2 transcript profiles in different individuals. Primer pair ASGR2RTF3 + ASGR2RTR2 gave RT-PCR products of different length (arrowed) according to the ASGR2 transcripts present. RNA extracted from the M + L cell fraction of 6 different individuals (A–F) gave either a profile consistent with transcripts encoding all isoforms (a, b, c, and d) or just with isoforms b and d. The liver cell line HepG2 showed all transcripts, the monocyte cell line THP1 showed only transcripts encoding isoforms b and d. The analysis was done using polyacrylamide gel electrophoresis. DNA size reference was pBR322/ Msp I (“Marker”, 238 bp and 120 bp sizes indicated). In addition to the expected products, additional major bands were visualized on polyacrylamide gel electrophoresis of the RT-PCR screen for ASGR2 transcripts (bracketed bands). It was considered possible that these represented heteroduplexes arising by cross-hybridization of complementary strands of the closely homologous true PCR products. To test this, each candidate heteroduplex band was excised from the gel, eluted into water, reamplified using the primers used in the initial PCR and the products electrophoresed on polyacrylamide. The results confirmed that the additional bands were hybrid duplexes containing complementary strands from nonidentical true PCR products: each hybrid yielded products consistent with amplification of two different template strands (data not shown).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Polyacrylamide Gel Electrophoresis, Hybridization, Polymerase Chain Reaction, Amplification

    Real-time PCR analyses. (a) Agarose gel electrophoresis of products from optimized real-time PCRs for ASGR1 (113 bp, primers ASG1RTF + ASG1RTR), ASGR2 (171 bp, primers ASG2RTF + ASG2RTR), and B2M (231 bp, primers B2MRTF + B2MRTR). Substrate was HepG2 cDNA. (b) Melting curve analysis of real-time PCR products from panel (a). The Tm for each product is indicated. (c and d) Relative quantification of ASGR1 (c) and ASGR2 (d) transcripts in cell-sorted monocytes (MC) compared with HepG2 (G2). B2M was used as the reference. (i) Fluorescence profiles obtained during real-time PCR. Samples were analyzed in duplicate, for clarity only one of each duplicate is shown. (ii) Melting curve analysis for the real-time PCR products in (i). (iii) Agarose gel electrophoresis of real-time PCR products (duplicate analyses shown). Neg indicates real-time PCR control containing water instead of nucleic acid template. In panels a, c(iii), and d(iii), M denotes pBR322/ Msp I size standard (238 bp and 120 bp sizes indicated).
    Figure Legend Snippet: Real-time PCR analyses. (a) Agarose gel electrophoresis of products from optimized real-time PCRs for ASGR1 (113 bp, primers ASG1RTF + ASG1RTR), ASGR2 (171 bp, primers ASG2RTF + ASG2RTR), and B2M (231 bp, primers B2MRTF + B2MRTR). Substrate was HepG2 cDNA. (b) Melting curve analysis of real-time PCR products from panel (a). The Tm for each product is indicated. (c and d) Relative quantification of ASGR1 (c) and ASGR2 (d) transcripts in cell-sorted monocytes (MC) compared with HepG2 (G2). B2M was used as the reference. (i) Fluorescence profiles obtained during real-time PCR. Samples were analyzed in duplicate, for clarity only one of each duplicate is shown. (ii) Melting curve analysis for the real-time PCR products in (i). (iii) Agarose gel electrophoresis of real-time PCR products (duplicate analyses shown). Neg indicates real-time PCR control containing water instead of nucleic acid template. In panels a, c(iii), and d(iii), M denotes pBR322/ Msp I size standard (238 bp and 120 bp sizes indicated).

    Techniques Used: Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Fluorescence

    Detection of ASGR1 and ASGR2 transcripts in peripheral blood mononuclear cells and localization to monocytes. Transcripts were detected using RT-PCR, the presence of ASGR1 transcripts was indicated by a 113 bp amplification product (primers ASG1RTF + ASG1RTR) and ASGR2 transcripts by a 171 bp product (primers ASG2RTF + ASG2RTR). HepG2 was used as a positive control and water as a negative control. Where necessary, β -2-microglobulin ( B2M ) was used as a reference to demonstrate equivalent amplification for each RNA preparation, (primers B2MRTF + B2MRTR, product 231 bp). (a) Analysis of RNA from monocyte cell line THP1 and PBMCs from two unrelated individuals (1 and 2). ASGR1 and ASGR2 transcripts were detected in all cases. (b) Analysis of RNA from peripheral blood cell fractions: monocytes + lymphocytes (M + L) and granulocytes + lymphocytes (G + L). ASGR1 and ASGR2 transcripts were detected in M + L but not in G + L. (c) Analysis of RNA from cell-sorted monocytes (mono, lanes 1–3 triplicate analyses) and lymphocytes (lymph, lanes 1–3 triplicate analyses) (data for water control not shown for ASGR1 and ASGR2 ). The data were obtained from a single experiment involving several gels: these are indicated by separate gel windows. For each panel, analysis was done using agarose gel electrophoresis with pBR322/ Msp I size standard (“Marker”, 238 bp and 120 bp indicated).
    Figure Legend Snippet: Detection of ASGR1 and ASGR2 transcripts in peripheral blood mononuclear cells and localization to monocytes. Transcripts were detected using RT-PCR, the presence of ASGR1 transcripts was indicated by a 113 bp amplification product (primers ASG1RTF + ASG1RTR) and ASGR2 transcripts by a 171 bp product (primers ASG2RTF + ASG2RTR). HepG2 was used as a positive control and water as a negative control. Where necessary, β -2-microglobulin ( B2M ) was used as a reference to demonstrate equivalent amplification for each RNA preparation, (primers B2MRTF + B2MRTR, product 231 bp). (a) Analysis of RNA from monocyte cell line THP1 and PBMCs from two unrelated individuals (1 and 2). ASGR1 and ASGR2 transcripts were detected in all cases. (b) Analysis of RNA from peripheral blood cell fractions: monocytes + lymphocytes (M + L) and granulocytes + lymphocytes (G + L). ASGR1 and ASGR2 transcripts were detected in M + L but not in G + L. (c) Analysis of RNA from cell-sorted monocytes (mono, lanes 1–3 triplicate analyses) and lymphocytes (lymph, lanes 1–3 triplicate analyses) (data for water control not shown for ASGR1 and ASGR2 ). The data were obtained from a single experiment involving several gels: these are indicated by separate gel windows. For each panel, analysis was done using agarose gel electrophoresis with pBR322/ Msp I size standard (“Marker”, 238 bp and 120 bp indicated).

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control, Negative Control, Agarose Gel Electrophoresis

    Related Articles

    RNA Sequencing Assay:

    Article Title: In vitro approaches to analysis of transcription termination
    Article Snippet: .. As a size marker, we use pBR322 DNA digested with Msp I (NEB) and end-labeled with γ-[32 P]ATP (3000 Ci/mmol, Perkin Elmer, cat #BLU-002A) and T4 polynucleotide kinase (NEB), or the RNA sequencing ladders generated as in 2.6; the former is sufficient for many applications ( ). .. Transfer the gel to a drying paper (such as blotting paper from Life Science Products, # LS238-3543), cover with plastic (any food wrap), and use a standard set up (such as BioRad GelDryer 583 and HydroTech vacuum pump); it takes ∼1 hour at 80°C to completely dry a full-size sequencing gel.

    Purification:

    Article Title: Locus-specific control of the de novo DNA methylation pathway in Arabidopsis by the CLASSY family
    Article Snippet: .. The final library products were further purified using an 8% polyacrylamide gel to excise 130–160nt products relative to the pBR322 DNA-MspI Digest ladder (New England Biolabs, Cat# E7323AA). .. The libraries were pooled and sequenced (single end 50bp, SE50) on a HiSeq 2500 machine (Illumina).

    Generated:

    Article Title: In vitro approaches to analysis of transcription termination
    Article Snippet: .. As a size marker, we use pBR322 DNA digested with Msp I (NEB) and end-labeled with γ-[32 P]ATP (3000 Ci/mmol, Perkin Elmer, cat #BLU-002A) and T4 polynucleotide kinase (NEB), or the RNA sequencing ladders generated as in 2.6; the former is sufficient for many applications ( ). .. Transfer the gel to a drying paper (such as blotting paper from Life Science Products, # LS238-3543), cover with plastic (any food wrap), and use a standard set up (such as BioRad GelDryer 583 and HydroTech vacuum pump); it takes ∼1 hour at 80°C to completely dry a full-size sequencing gel.

    other:

    Article Title: Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization
    Article Snippet: Reagents included casein (#C-5890, Sigma); pBR322 Msp I digest (#303-2S, New England Biolabs) and Bst NI digests (#301-4S, New England Biolabs); M13 DNA (#770704, United States Biochemical); control DNA (G304A 5491901, Promega); W1282X carrier DNA (NAO454OA, Coriell Cell Repository, Camden, NJ); and Immobilon-S (IMS02, Millipore).

    Marker:

    Article Title: In vitro approaches to analysis of transcription termination
    Article Snippet: .. As a size marker, we use pBR322 DNA digested with Msp I (NEB) and end-labeled with γ-[32 P]ATP (3000 Ci/mmol, Perkin Elmer, cat #BLU-002A) and T4 polynucleotide kinase (NEB), or the RNA sequencing ladders generated as in 2.6; the former is sufficient for many applications ( ). .. Transfer the gel to a drying paper (such as blotting paper from Life Science Products, # LS238-3543), cover with plastic (any food wrap), and use a standard set up (such as BioRad GelDryer 583 and HydroTech vacuum pump); it takes ∼1 hour at 80°C to completely dry a full-size sequencing gel.

    Article Title: NeuroD1 Gene and Interleukin-18 Gene Polymorphisms in Type 1 Diabetes in the Dalmatian Population of Southern Croatia
    Article Snippet: .. For determination of fragment length DNA marker, pBR322 DNA-MspI Digest (New England BioLabs) was used. .. IL-18 polymorphisms were detected by using PCR sequence-specific primers, according to the previous reports ( , ).

    Article Title: Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor ?1 subunit
    Article Snippet: .. The molecular weight marker was the pBR322 DNA– Msp I digest (New England Biolabs, Ipswich, MA, USA). ..

    Staining:

    Article Title: Fetal Mouse Cyp1b1 and Transplacental Carcinogenesis from Maternal Exposure to Dibenzo[a,l]pyrene
    Article Snippet: .. A molecular weight ladder of Msp I cut pBR322 DNA (New England Biolabs, Ipswich, MA) and ethidium bromide (Sigma Chemical Co., St. Louis, MO) was used to stain the DNA, followed by UV visualization. .. Litter sizes per dam were compared between treatments using the exact Kruskal-Wallis test (SAS 9.1.3 Npar1way procedure).

    Molecular Weight:

    Article Title: Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor ?1 subunit
    Article Snippet: .. The molecular weight marker was the pBR322 DNA– Msp I digest (New England Biolabs, Ipswich, MA, USA). ..

    Article Title: Fetal Mouse Cyp1b1 and Transplacental Carcinogenesis from Maternal Exposure to Dibenzo[a,l]pyrene
    Article Snippet: .. A molecular weight ladder of Msp I cut pBR322 DNA (New England Biolabs, Ipswich, MA) and ethidium bromide (Sigma Chemical Co., St. Louis, MO) was used to stain the DNA, followed by UV visualization. .. Litter sizes per dam were compared between treatments using the exact Kruskal-Wallis test (SAS 9.1.3 Npar1way procedure).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    New England Biolabs pbr322 dna msp
    Cells with fast initial decay kinetics express β 1 subunit mRNA A and B , scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B , represents a recording dominated by brief and large IPSCs ( ). C , ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A 1 value than A 2 whereas the opposite is true for the slower cell. D , ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β 1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the <t>Msp</t> I digest of <t>pBR322.</t> The size of each fragment is noted on the right. The parameters for β 1 negative cells in lane 4: A 1 = 9.80, τ 1 = 13.75, A 2 = 14.17, τ 2 = 64.61, τ DW = 43.82l; and lane 5: A 1 = 12.94, τ 1 = 12.94, A 2 = 5.69, τ 2 = 77.21, τ DW = 29.63. The β 1 positive cell in lane 6: A 1 = 18.05, τ 1 = 15.11, A 2 = 4.15, τ 2 = 74.44, τ DW = 26.20.
    Pbr322 Dna Msp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbr322 dna msp/product/New England Biolabs
    Average 93 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    pbr322 dna msp - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    Image Search Results


    Cells with fast initial decay kinetics express β 1 subunit mRNA A and B , scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B , represents a recording dominated by brief and large IPSCs ( ). C , ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A 1 value than A 2 whereas the opposite is true for the slower cell. D , ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β 1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the Msp I digest of pBR322. The size of each fragment is noted on the right. The parameters for β 1 negative cells in lane 4: A 1 = 9.80, τ 1 = 13.75, A 2 = 14.17, τ 2 = 64.61, τ DW = 43.82l; and lane 5: A 1 = 12.94, τ 1 = 12.94, A 2 = 5.69, τ 2 = 77.21, τ DW = 29.63. The β 1 positive cell in lane 6: A 1 = 18.05, τ 1 = 15.11, A 2 = 4.15, τ 2 = 74.44, τ DW = 26.20.

    Journal: The Journal of Physiology

    Article Title: Fast IPSCs in rat thalamic reticular nucleus require the GABAA receptor ?1 subunit

    doi: 10.1113/jphysiol.2006.106617

    Figure Lengend Snippet: Cells with fast initial decay kinetics express β 1 subunit mRNA A and B , scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B , represents a recording dominated by brief and large IPSCs ( ). C , ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A 1 value than A 2 whereas the opposite is true for the slower cell. D , ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β 1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the Msp I digest of pBR322. The size of each fragment is noted on the right. The parameters for β 1 negative cells in lane 4: A 1 = 9.80, τ 1 = 13.75, A 2 = 14.17, τ 2 = 64.61, τ DW = 43.82l; and lane 5: A 1 = 12.94, τ 1 = 12.94, A 2 = 5.69, τ 2 = 77.21, τ DW = 29.63. The β 1 positive cell in lane 6: A 1 = 18.05, τ 1 = 15.11, A 2 = 4.15, τ 2 = 74.44, τ DW = 26.20.

    Article Snippet: The molecular weight marker was the pBR322 DNA– Msp I digest (New England Biolabs, Ipswich, MA, USA).

    Techniques: Isolation, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control, Polymerase Chain Reaction, Molecular Weight, Marker

    Pre-gel hybridization parameters. ( A ) Schematic diagram of pre-gel hybridization technique. ( B ) Comparison of 1 pmol each of biotinylated PNA and DNA pBR322 15-mer probes hybridized to 2 ng of Bst NI-digested pBR322. Lanes: 1, PNA probe; 2, DNA probe. ( C ) The effect of different sodium ion concentrations on a mixture of Msp I (50 ng) and Bst NI (15 ng) digests of pBR322. One picomole of fluorescently labeled PNA was used. Lanes: 1, no salt; 2, 0.1 mM; 3, 0.5 mM; 4, 1 mM; 5, 5 mM; 6, 10 mM; 7, 20 mM; 8, 30 mM; 9, 40 mM; 10, 50 mM; 11, 75 mM; 12, 100 mM; 13, 250 mM; and 14, 500 mM. ( D ) Size separation hybridizing 1 pmol lambda fluorescein-tagged PNA probe and 50 ng of each digest. Lanes: 1, Hin dIII/ Bam HI; 2, Eco RI/ Bam HI; 3, Bst EII; 4, Hin dIII.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization

    doi:

    Figure Lengend Snippet: Pre-gel hybridization parameters. ( A ) Schematic diagram of pre-gel hybridization technique. ( B ) Comparison of 1 pmol each of biotinylated PNA and DNA pBR322 15-mer probes hybridized to 2 ng of Bst NI-digested pBR322. Lanes: 1, PNA probe; 2, DNA probe. ( C ) The effect of different sodium ion concentrations on a mixture of Msp I (50 ng) and Bst NI (15 ng) digests of pBR322. One picomole of fluorescently labeled PNA was used. Lanes: 1, no salt; 2, 0.1 mM; 3, 0.5 mM; 4, 1 mM; 5, 5 mM; 6, 10 mM; 7, 20 mM; 8, 30 mM; 9, 40 mM; 10, 50 mM; 11, 75 mM; 12, 100 mM; 13, 250 mM; and 14, 500 mM. ( D ) Size separation hybridizing 1 pmol lambda fluorescein-tagged PNA probe and 50 ng of each digest. Lanes: 1, Hin dIII/ Bam HI; 2, Eco RI/ Bam HI; 3, Bst EII; 4, Hin dIII.

    Article Snippet: Reagents included casein (#C-5890, Sigma); pBR322 Msp I digest (#303-2S, New England Biolabs) and Bst NI digests (#301-4S, New England Biolabs); M13 DNA (#770704, United States Biochemical); control DNA (G304A 5491901, Promega); W1282X carrier DNA (NAO454OA, Coriell Cell Repository, Camden, NJ); and Immobilon-S (IMS02, Millipore).

    Techniques: Hybridization, Labeling

    CE separation of a mixed Msp I- and Bst NI-digested pBR322. ( A ) UV detection. Voltage: 5 kV. Detection: 260 nm. Buffer: TBE/7 M urea/0.5% PEO (8 MDa). ( B ) LIF detection. Voltage: 5 kV. Detection: LIF 488 nm/520 nm. Buffer: 1× TBE/1% PEO (4 MDa). Temperature 50°C.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization

    doi:

    Figure Lengend Snippet: CE separation of a mixed Msp I- and Bst NI-digested pBR322. ( A ) UV detection. Voltage: 5 kV. Detection: 260 nm. Buffer: TBE/7 M urea/0.5% PEO (8 MDa). ( B ) LIF detection. Voltage: 5 kV. Detection: LIF 488 nm/520 nm. Buffer: 1× TBE/1% PEO (4 MDa). Temperature 50°C.

    Article Snippet: Reagents included casein (#C-5890, Sigma); pBR322 Msp I digest (#303-2S, New England Biolabs) and Bst NI digests (#301-4S, New England Biolabs); M13 DNA (#770704, United States Biochemical); control DNA (G304A 5491901, Promega); W1282X carrier DNA (NAO454OA, Coriell Cell Repository, Camden, NJ); and Immobilon-S (IMS02, Millipore).

    Techniques: Multiple Displacement Amplification

    ASGR1 transcript profiles in different individuals. Nested PCR was used to detect ASGR1 transcripts T1 and T2 (first round primers A1CDSF1 + A1CDSR1; second round A1TEST2 + ASG1RTR). A product of 408 bp represented T1, present in RNA from the M + L cell fraction of five individuals (A–E) and also in HepG2 and THP1; a 291 bp product indicated the presence of T2 (individual E, and, very faintly, in HepG2). The RT-PCR product for individual A was coloaded with the DNA size standard (pBR322/ Msp I, 238 bp and 120 bp sizes indicated). Analysis was done using agarose gel electrophoresis. The figure is a composite of various data (indicated by individual gel windows).

    Journal: Molecular Biology International

    Article Title: ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    doi: 10.1155/2012/283974

    Figure Lengend Snippet: ASGR1 transcript profiles in different individuals. Nested PCR was used to detect ASGR1 transcripts T1 and T2 (first round primers A1CDSF1 + A1CDSR1; second round A1TEST2 + ASG1RTR). A product of 408 bp represented T1, present in RNA from the M + L cell fraction of five individuals (A–E) and also in HepG2 and THP1; a 291 bp product indicated the presence of T2 (individual E, and, very faintly, in HepG2). The RT-PCR product for individual A was coloaded with the DNA size standard (pBR322/ Msp I, 238 bp and 120 bp sizes indicated). Analysis was done using agarose gel electrophoresis. The figure is a composite of various data (indicated by individual gel windows).

    Article Snippet: The DNA size standard pBR322/Msp I (New England Biolabs, Hertfordshire, UK) was used for all electrophoretic analyses.

    Techniques: Nested PCR, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis

    ASGR2 transcript profiles in different individuals. Primer pair ASGR2RTF3 + ASGR2RTR2 gave RT-PCR products of different length (arrowed) according to the ASGR2 transcripts present. RNA extracted from the M + L cell fraction of 6 different individuals (A–F) gave either a profile consistent with transcripts encoding all isoforms (a, b, c, and d) or just with isoforms b and d. The liver cell line HepG2 showed all transcripts, the monocyte cell line THP1 showed only transcripts encoding isoforms b and d. The analysis was done using polyacrylamide gel electrophoresis. DNA size reference was pBR322/ Msp I (“Marker”, 238 bp and 120 bp sizes indicated). In addition to the expected products, additional major bands were visualized on polyacrylamide gel electrophoresis of the RT-PCR screen for ASGR2 transcripts (bracketed bands). It was considered possible that these represented heteroduplexes arising by cross-hybridization of complementary strands of the closely homologous true PCR products. To test this, each candidate heteroduplex band was excised from the gel, eluted into water, reamplified using the primers used in the initial PCR and the products electrophoresed on polyacrylamide. The results confirmed that the additional bands were hybrid duplexes containing complementary strands from nonidentical true PCR products: each hybrid yielded products consistent with amplification of two different template strands (data not shown).

    Journal: Molecular Biology International

    Article Title: ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    doi: 10.1155/2012/283974

    Figure Lengend Snippet: ASGR2 transcript profiles in different individuals. Primer pair ASGR2RTF3 + ASGR2RTR2 gave RT-PCR products of different length (arrowed) according to the ASGR2 transcripts present. RNA extracted from the M + L cell fraction of 6 different individuals (A–F) gave either a profile consistent with transcripts encoding all isoforms (a, b, c, and d) or just with isoforms b and d. The liver cell line HepG2 showed all transcripts, the monocyte cell line THP1 showed only transcripts encoding isoforms b and d. The analysis was done using polyacrylamide gel electrophoresis. DNA size reference was pBR322/ Msp I (“Marker”, 238 bp and 120 bp sizes indicated). In addition to the expected products, additional major bands were visualized on polyacrylamide gel electrophoresis of the RT-PCR screen for ASGR2 transcripts (bracketed bands). It was considered possible that these represented heteroduplexes arising by cross-hybridization of complementary strands of the closely homologous true PCR products. To test this, each candidate heteroduplex band was excised from the gel, eluted into water, reamplified using the primers used in the initial PCR and the products electrophoresed on polyacrylamide. The results confirmed that the additional bands were hybrid duplexes containing complementary strands from nonidentical true PCR products: each hybrid yielded products consistent with amplification of two different template strands (data not shown).

    Article Snippet: The DNA size standard pBR322/Msp I (New England Biolabs, Hertfordshire, UK) was used for all electrophoretic analyses.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polyacrylamide Gel Electrophoresis, Hybridization, Polymerase Chain Reaction, Amplification

    Real-time PCR analyses. (a) Agarose gel electrophoresis of products from optimized real-time PCRs for ASGR1 (113 bp, primers ASG1RTF + ASG1RTR), ASGR2 (171 bp, primers ASG2RTF + ASG2RTR), and B2M (231 bp, primers B2MRTF + B2MRTR). Substrate was HepG2 cDNA. (b) Melting curve analysis of real-time PCR products from panel (a). The Tm for each product is indicated. (c and d) Relative quantification of ASGR1 (c) and ASGR2 (d) transcripts in cell-sorted monocytes (MC) compared with HepG2 (G2). B2M was used as the reference. (i) Fluorescence profiles obtained during real-time PCR. Samples were analyzed in duplicate, for clarity only one of each duplicate is shown. (ii) Melting curve analysis for the real-time PCR products in (i). (iii) Agarose gel electrophoresis of real-time PCR products (duplicate analyses shown). Neg indicates real-time PCR control containing water instead of nucleic acid template. In panels a, c(iii), and d(iii), M denotes pBR322/ Msp I size standard (238 bp and 120 bp sizes indicated).

    Journal: Molecular Biology International

    Article Title: ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    doi: 10.1155/2012/283974

    Figure Lengend Snippet: Real-time PCR analyses. (a) Agarose gel electrophoresis of products from optimized real-time PCRs for ASGR1 (113 bp, primers ASG1RTF + ASG1RTR), ASGR2 (171 bp, primers ASG2RTF + ASG2RTR), and B2M (231 bp, primers B2MRTF + B2MRTR). Substrate was HepG2 cDNA. (b) Melting curve analysis of real-time PCR products from panel (a). The Tm for each product is indicated. (c and d) Relative quantification of ASGR1 (c) and ASGR2 (d) transcripts in cell-sorted monocytes (MC) compared with HepG2 (G2). B2M was used as the reference. (i) Fluorescence profiles obtained during real-time PCR. Samples were analyzed in duplicate, for clarity only one of each duplicate is shown. (ii) Melting curve analysis for the real-time PCR products in (i). (iii) Agarose gel electrophoresis of real-time PCR products (duplicate analyses shown). Neg indicates real-time PCR control containing water instead of nucleic acid template. In panels a, c(iii), and d(iii), M denotes pBR322/ Msp I size standard (238 bp and 120 bp sizes indicated).

    Article Snippet: The DNA size standard pBR322/Msp I (New England Biolabs, Hertfordshire, UK) was used for all electrophoretic analyses.

    Techniques: Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Fluorescence

    Detection of ASGR1 and ASGR2 transcripts in peripheral blood mononuclear cells and localization to monocytes. Transcripts were detected using RT-PCR, the presence of ASGR1 transcripts was indicated by a 113 bp amplification product (primers ASG1RTF + ASG1RTR) and ASGR2 transcripts by a 171 bp product (primers ASG2RTF + ASG2RTR). HepG2 was used as a positive control and water as a negative control. Where necessary, β -2-microglobulin ( B2M ) was used as a reference to demonstrate equivalent amplification for each RNA preparation, (primers B2MRTF + B2MRTR, product 231 bp). (a) Analysis of RNA from monocyte cell line THP1 and PBMCs from two unrelated individuals (1 and 2). ASGR1 and ASGR2 transcripts were detected in all cases. (b) Analysis of RNA from peripheral blood cell fractions: monocytes + lymphocytes (M + L) and granulocytes + lymphocytes (G + L). ASGR1 and ASGR2 transcripts were detected in M + L but not in G + L. (c) Analysis of RNA from cell-sorted monocytes (mono, lanes 1–3 triplicate analyses) and lymphocytes (lymph, lanes 1–3 triplicate analyses) (data for water control not shown for ASGR1 and ASGR2 ). The data were obtained from a single experiment involving several gels: these are indicated by separate gel windows. For each panel, analysis was done using agarose gel electrophoresis with pBR322/ Msp I size standard (“Marker”, 238 bp and 120 bp indicated).

    Journal: Molecular Biology International

    Article Title: ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    doi: 10.1155/2012/283974

    Figure Lengend Snippet: Detection of ASGR1 and ASGR2 transcripts in peripheral blood mononuclear cells and localization to monocytes. Transcripts were detected using RT-PCR, the presence of ASGR1 transcripts was indicated by a 113 bp amplification product (primers ASG1RTF + ASG1RTR) and ASGR2 transcripts by a 171 bp product (primers ASG2RTF + ASG2RTR). HepG2 was used as a positive control and water as a negative control. Where necessary, β -2-microglobulin ( B2M ) was used as a reference to demonstrate equivalent amplification for each RNA preparation, (primers B2MRTF + B2MRTR, product 231 bp). (a) Analysis of RNA from monocyte cell line THP1 and PBMCs from two unrelated individuals (1 and 2). ASGR1 and ASGR2 transcripts were detected in all cases. (b) Analysis of RNA from peripheral blood cell fractions: monocytes + lymphocytes (M + L) and granulocytes + lymphocytes (G + L). ASGR1 and ASGR2 transcripts were detected in M + L but not in G + L. (c) Analysis of RNA from cell-sorted monocytes (mono, lanes 1–3 triplicate analyses) and lymphocytes (lymph, lanes 1–3 triplicate analyses) (data for water control not shown for ASGR1 and ASGR2 ). The data were obtained from a single experiment involving several gels: these are indicated by separate gel windows. For each panel, analysis was done using agarose gel electrophoresis with pBR322/ Msp I size standard (“Marker”, 238 bp and 120 bp indicated).

    Article Snippet: The DNA size standard pBR322/Msp I (New England Biolabs, Hertfordshire, UK) was used for all electrophoretic analyses.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control, Negative Control, Agarose Gel Electrophoresis