Real time RT-PCR with a newly designed set of primers confirmed the presence of 2b and 2x/d myosin heavy chain mRNAs in the rat slow soleus muscle
Language English Country Czech Republic Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
19154088
DOI
10.33549/physiolres.931676
PII: 1676
Knihovny.cz E-resources
- MeSH
- DNA Primers * MeSH
- Muscle, Skeletal chemistry MeSH
- Rats MeSH
- RNA, Messenger analysis MeSH
- Polymerase Chain Reaction * MeSH
- Rats, Inbred Lew MeSH
- Protein Isoforms MeSH
- Muscle Fibers, Slow-Twitch chemistry MeSH
- Muscle Fibers, Fast-Twitch chemistry MeSH
- Myosin Heavy Chains genetics MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA Primers * MeSH
- RNA, Messenger MeSH
- Protein Isoforms MeSH
- Myosin Heavy Chains MeSH
In order to re-evaluate the presence and relative quantity of 2b and 2x/d myosin heavy chain (MyHC) transcripts in rat slow soleus muscle by using real time RT-PCR we have compared the available relevant cDNA sequences and designed a new set of primers having similar melting temperatures, matching separate MyHC exons in the regions of maximal differences in MyHC coding sequences, and containing G or C at the 3 -end. These also yielded PCR products of corresponding length, which is an important requirement for real time RT-PCR quantification. The experiments were performed on 8-month-old inbred female Lewis strain rats used in our current study of regenerating transplanted muscles. The real time RT-PCR measurement confirmed the expression of all four MyHC mRNAs (type 1, 2a, 2x/d and 2b) in both fast extensor digitorum longus and slow soleus muscles, although in the soleus muscle of adult rats, only type 1 and 2a protein isoforms can be usually detected.
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