View in EDS

Analyzing Exonuclease-Induced Hyperchromicity by UV Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment.

Saved in:
Bibliographic Details
Title: Analyzing Exonuclease-Induced Hyperchromicity by UV Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment.
Authors: Ackerman, Megan M.1, Ricciardi, Christopher1, Weiss, David1, Chant, Alan2, Kraemer-Chant, Christina M.1 cchant@smcvt.edu
Source: Journal of Chemical Education. Dec2016, Vol. 93 Issue 12, p2089-2095. 7p.
Subjects: Exonucleases, Hyperchromic effect, Spectrometry, Biochemistry experiments, Undergraduates
Abstract: An undergraduate biochemistry laboratory experiment is described that utilizes free online bioinformatics tools along with readily available exonucleases to study the effects of base stacking and hydrogen bonding on the UV absorbance of DNA samples. UV absorbance of double-stranded DNA at the λ[submax] is decreased when the DNA bases are involved in hydrogen bonding and formation of secondary structure. When an exonuclease is added to the solution containing the DNA, the strand is digested and the interactions disappear, leading to an increase in the absorbance called hyperchromicity. This experiment utilizes exonuclease digestion of DNA to show students how base interactions and secondary structure can alter the spectroscopic properties of a sample and, by extension, how apparent concentration as calculated with Beer-Lambert's law is not necessarily representative of the true concentration of DNA in solution. Teaching applications of this laboratory experiment include enzyme kinetics and activity, secondary structure of single-stranded DNA (and its parallel to RNA structure), and an introduction to bioinformatics. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Chemical Education is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
FullText Text:
  Availability: 0
Header DbId: egs
DbLabel: Engineering Source
An: 120234207
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
IllustrationInfo
Items – Name: Title
  Label: Title
  Group: Ti
  Data: Analyzing Exonuclease-Induced Hyperchromicity by UV Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment.
– Name: Author
  Label: Authors
  Group: Au
  Data: <searchLink fieldCode="AR" term="%22Ackerman%2C+Megan+M%2E%22">Ackerman, Megan M.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Ricciardi%2C+Christopher%22">Ricciardi, Christopher</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Weiss%2C+David%22">Weiss, David</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Chant%2C+Alan%22">Chant, Alan</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Kraemer-Chant%2C+Christina+M%2E%22">Kraemer-Chant, Christina M.</searchLink><relatesTo>1</relatesTo><i> cchant@smcvt.edu</i>
– Name: TitleSource
  Label: Source
  Group: Src
  Data: <searchLink fieldCode="JN" term="%22Journal+of+Chemical+Education%22">Journal of Chemical Education</searchLink>. Dec2016, Vol. 93 Issue 12, p2089-2095. 7p.
– Name: Subject
  Label: Subjects
  Group: Su
  Data: <searchLink fieldCode="DE" term="%22Exonucleases%22">Exonucleases</searchLink><br /><searchLink fieldCode="DE" term="%22Hyperchromic+effect%22">Hyperchromic effect</searchLink><br /><searchLink fieldCode="DE" term="%22Spectrometry%22">Spectrometry</searchLink><br /><searchLink fieldCode="DE" term="%22Biochemistry+experiments%22">Biochemistry experiments</searchLink><br /><searchLink fieldCode="DE" term="%22Undergraduates%22">Undergraduates</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: An undergraduate biochemistry laboratory experiment is described that utilizes free online bioinformatics tools along with readily available exonucleases to study the effects of base stacking and hydrogen bonding on the UV absorbance of DNA samples. UV absorbance of double-stranded DNA at the λ[submax] is decreased when the DNA bases are involved in hydrogen bonding and formation of secondary structure. When an exonuclease is added to the solution containing the DNA, the strand is digested and the interactions disappear, leading to an increase in the absorbance called hyperchromicity. This experiment utilizes exonuclease digestion of DNA to show students how base interactions and secondary structure can alter the spectroscopic properties of a sample and, by extension, how apparent concentration as calculated with Beer-Lambert's law is not necessarily representative of the true concentration of DNA in solution. Teaching applications of this laboratory experiment include enzyme kinetics and activity, secondary structure of single-stranded DNA (and its parallel to RNA structure), and an introduction to bioinformatics. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Journal of Chemical Education is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=egs&AN=120234207
RecordInfo BibRecord:
  BibEntity:
    Identifiers:
      – Type: doi
        Value: 10.1021/acs.jchemed.6b00095
    Languages:
      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        PageCount: 7
        StartPage: 2089
    Subjects:
      – SubjectFull: Exonucleases
        Type: general
      – SubjectFull: Hyperchromic effect
        Type: general
      – SubjectFull: Spectrometry
        Type: general
      – SubjectFull: Biochemistry experiments
        Type: general
      – SubjectFull: Undergraduates
        Type: general
    Titles:
      – TitleFull: Analyzing Exonuclease-Induced Hyperchromicity by UV Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment.
        Type: main
  BibRelationships:
    HasContributorRelationships:
      – PersonEntity:
          Name:
            NameFull: Ackerman, Megan M.
      – PersonEntity:
          Name:
            NameFull: Ricciardi, Christopher
      – PersonEntity:
          Name:
            NameFull: Weiss, David
      – PersonEntity:
          Name:
            NameFull: Chant, Alan
      – PersonEntity:
          Name:
            NameFull: Kraemer-Chant, Christina M.
    IsPartOfRelationships:
      – BibEntity:
          Dates:
            – D: 13
              M: 12
              Text: Dec2016
              Type: published
              Y: 2016
          Identifiers:
            – Type: issn-print
              Value: 00219584
          Numbering:
            – Type: volume
              Value: 93
            – Type: issue
              Value: 12
          Titles:
            – TitleFull: Journal of Chemical Education
              Type: main
ResultId 1