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Absolute Quantification of RXRG isoforms
Author(s) -
Skotarczak Teagan,
Karnath Erin,
Monin Lydia,
Shimanovich Yana,
Bistulfi Gaia
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.908.24
Subject(s) - gene isoform , amplicon , biology , gene , alternative splicing , terminator (solar) , genetics , computational biology , transcription (linguistics) , microbiology and biotechnology , polymerase chain reaction , ionosphere , linguistics , philosophy , physics , astronomy
Many genes are transcribed as multiple variants known as isoforms. Isoforms are transcripts from the same locus with either a different transcription start site or alternative splicing. Isoforms might be characterized by different functions and/or stability. Different isoforms of the same gene can vary in length from a few bases to several kilobases. In some cases, there is no region of the transcript specific to only one isoform, making quantification problematic. We compared the isoforms' sequences using bioinformatic tools freely available online. We utilized a one‐step real‐time RT‐PCR protocol, using primers specific to one or more isoforms, which included a T7 promoter and terminator sequences. After confirming amplicon identity by DNA sequencing, we built RNA standard curves. Analysis of the data shows acceptable specificity and efficiency for RXRG3 primers as well as primers picking up all three isoforms. We are currently in the process of assessing primers for RXRG1 and RXRG3. Support or Funding Information This work was supported by funding from D'Youville College, Buffalo NY