Increased yield of PCR products by addition of T4 gene 32 protein to the SMART (TM) PCR cDNA synthesis system

Citation
C. Villalva et al., Increased yield of PCR products by addition of T4 gene 32 protein to the SMART (TM) PCR cDNA synthesis system, BIOTECHNIQU, 31(1), 2001, pp. 81
Citations number
12
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOTECHNIQUES
ISSN journal
07366205 → ACNP
Volume
31
Issue
1
Year of publication
2001
Database
ISI
SICI code
0736-6205(200107)31:1<81:IYOPPB>2.0.ZU;2-M
Abstract
Under certain conditions, T4 gene 32 protein is known to increase the effic iency of different enzymes, such as Tag DNA polymerase, reverse transcripta se, and telomerase. In this study We compared the efficiency of the SMART ( TM) PCR cDNA synthesis kit with and without the T4 gene 32 protein. The use of this cDNA synthesis procedure, in combination with T4 gene 32 protein, increases the yield of RT-PCR products from approximately 90% to 150%. This effect is even observed for long mRNA templates and low concentrations of total RNA (25 ng). Therefore, we suggest the addition of T4 gene 32 protein in the RT-PCR mixture to increase the efficiency of cDNA synthesis, partic ularly in cases when low amounts of tissue are used.