Description
General description
Procedures are provided for one-step and two-step RT-PCR reactions.
One-step: In a single tube, eAMV™ RT and AccuTaq LA act sequentially to first produce cDNA and then immediately amplify by PCR. This provides quick, sensitive analysis of RNA.
Two-step: Each reaction is individually optimized for greater yields with high fidelity, when protocol requires multiple amplifications, or if maximum yield is more important than maximum convenience.
Application
Enhanced Avian HS RT-PCR Kit has been used to synthesize the cDNA first-strand during reverse transcriptase PCR (RT-PCR) analysis.
Features and Benefits
Greater transcription lengths than other reverse transcriptases, generating cDNA up to 14.1 Kb.
Higher sensitivity for detecting low abundance messages. eAMV™ RT is able to transcribe RNA that other reverse transcriptases cannot detect.
Unsurpassed transcription through difficult secondary structure.
Increased sensitivity, specificity and yield from JumpStart AccuTaq LA DNA Polymerase.
Other Notes
Reverse Transcriptase PCR (RT-PCR) is a powerful tool used to study gene expression. The Enhanced Avian HS RT-PCR kit utilizes an enhanced avian myeloblastosis virus reverse transcriptase (eAMV-RT) enzyme that offers superior performance in comparison to standard AMV-RT and Moloney murine leukemia virus reverse transcriptase (MMLV-RT). eAMV RT is an exceptionally robust enzyme with an enhanced ability to transcribe through difficult secondary structure at elevated temperatures (up to 65 °C) making it the ideal enzyme for producing high quality full-length cDNA (up to 14.1 kb) from total RNA or poly(A)⁺ RNA. JumpStart™ AccuTaq™ LA DNA polymerase mix is also provided to eliminate non-specific amplification and increase specificity and sensitivity. The combination of these two enzymes provides a quality system that offers the versatility of a one-step or two-step RT-PCR protocol.
Legal Information
No license is conveyed with the purchase of this product under any of US Patents Nos. 5,804,375, 5,994,056, 6,171,785, 6,214,979, 5,538,848, 5,723,591, 5,876,930, 6,030,787, and 6,258,569, and corresponding patents outside the United States, or any other patents or patent applications, relating to the 5′ Nuclease and dsDNA-Binding Dye Processes. For further information contact the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
AccuTaq is a trademark of Sigma-Aldrich Co. LLC
JumpStart is a trademark of Sigma-Aldrich Co. LLC
eAMV is a trademark of Sigma-Aldrich Co. LLC
