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Quantitation of Small RNAs in Caenorhabditis elegans Using Real-Time RT-PCR

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003914/00001

Material Information

Title: Quantitation of Small RNAs in Caenorhabditis elegans Using Real-Time RT-PCR
Physical Description: Book
Language: English
Creator: Felsen, Matthew H.
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2008
Publication Date: 2008

Subjects

Subjects / Keywords: Real-Time PCR
RNA
PTGS
RT-PCR
Nematodes
Worms
C. elegans
Small RNAs
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: RNAi is a gene-regulatory process mediated by small RNAs approximately 22 nucleotides in length and is conserved across a wide variety of organisms, including worms, flies, and humans. Three proteins, rha-1, rde-2, and mut-7, are involved in RNAi and silencing of repetitive transgene arrays in the germline, and mutant strains show defects in these processes. In order to examine the possibility that small RNAs are abnormally expressed in these worms and in double mutant strains, a modified real-time RT-PCR procedure was employed to quantitate the expression of three distinct classes of small RNA molecules: microRNAs, tiny non-coding RNAs, and X cluster RNAs. Results show that the rha-1(tm329) strain overexpresses all RNAs tested, while the rde-2(ne221) and mut-7(pk204) strains have reduced expression in the tncRNAs and X cluster RNAs, but not miRNAs. Further, the introduction of the rha-1(tm329) mutation into the rde-2 and mut-7 strains rescues expression to near wild-type levels of many of the small RNAs in the rde-2;rha-1 and mut-7;rha-1 double mutant strains. The mechanism for this rescue is currently unknown.
Statement of Responsibility: by Matthew H. Felsen
Thesis: Thesis (B.A.) -- New College of Florida, 2008
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Walstrom, Katherine

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2008 F3
System ID: NCFE003914:00001

Permanent Link: http://ncf.sobek.ufl.edu/NCFE003914/00001

Material Information

Title: Quantitation of Small RNAs in Caenorhabditis elegans Using Real-Time RT-PCR
Physical Description: Book
Language: English
Creator: Felsen, Matthew H.
Publisher: New College of Florida
Place of Publication: Sarasota, Fla.
Creation Date: 2008
Publication Date: 2008

Subjects

Subjects / Keywords: Real-Time PCR
RNA
PTGS
RT-PCR
Nematodes
Worms
C. elegans
Small RNAs
Genre: bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: RNAi is a gene-regulatory process mediated by small RNAs approximately 22 nucleotides in length and is conserved across a wide variety of organisms, including worms, flies, and humans. Three proteins, rha-1, rde-2, and mut-7, are involved in RNAi and silencing of repetitive transgene arrays in the germline, and mutant strains show defects in these processes. In order to examine the possibility that small RNAs are abnormally expressed in these worms and in double mutant strains, a modified real-time RT-PCR procedure was employed to quantitate the expression of three distinct classes of small RNA molecules: microRNAs, tiny non-coding RNAs, and X cluster RNAs. Results show that the rha-1(tm329) strain overexpresses all RNAs tested, while the rde-2(ne221) and mut-7(pk204) strains have reduced expression in the tncRNAs and X cluster RNAs, but not miRNAs. Further, the introduction of the rha-1(tm329) mutation into the rde-2 and mut-7 strains rescues expression to near wild-type levels of many of the small RNAs in the rde-2;rha-1 and mut-7;rha-1 double mutant strains. The mechanism for this rescue is currently unknown.
Statement of Responsibility: by Matthew H. Felsen
Thesis: Thesis (B.A.) -- New College of Florida, 2008
Electronic Access: RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography: Includes bibliographical references.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Local: Faculty Sponsor: Walstrom, Katherine

Record Information

Source Institution: New College of Florida
Holding Location: New College of Florida
Rights Management: Applicable rights reserved.
Classification: local - S.T. 2008 F3
System ID: NCFE003914:00001

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