A4464
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 | mRNA from HEK293 cells was reverse transcribed using eAMV¿Reverse Transcriptase to a known length of 14.1 kb. The resulting cDNA was amplified using JumpStart REDTaq with PCR primer sets (RNA Inspector Kit, Product Code INSP-1) chosen at different distances from the poly (A)+ tail resulting in the following PCR products. Successful amplifications demonstrate the integrity of the cDNA up to 14 kb. Lanes 2-8 are primer sets for p619, Lane 10 is actin and Lane 11 is GAPDH. Lane 1 Marker Lane 2 908 bp PCR product, 2,180 transcript size Lane 3 796 bp PCR product, 5,760 transcript size Lane 4 708 bp PCR product, 6,875 transcript size Lane 5 608 bp PCR product, 9,970 transcript size Lane 6 499 bp PCR product, 10,980 transcript size Lane 7 396 bp PCR product, 12,890 transcript size Lane 8 300 bp PCR product, 14,150 transcript size Lane 9 Negative Control (No RT) Lane 10 349 bp PCR product, 1,060 transcript size Lane 11 452 bp PCR product, 880 transcript size |
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A4464
Sigma
Enhanced Avian Reverse Transcriptase [eAMV™ RT]
| MDL number: | MFCD01634820 |
Description
| Application | Enhanced Avian (eAMV) Reverse Transcriptase is a highly purified, avian myeloblastosis virus reverse transcriptase (AMV-RT) that offers superior performance in comparison to standard AMV-RT or standard Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV-RT). This exceptionally robust AMV-RT has an enhanced ability to detect low abundance messages, transcribe through difficult secondary structure at elevated temperatures (up to 65 °C), and transcribe mRNA templates up to 14.1 kb. |
| Features and Benefits | • Greater sensitivity for low abundance mRNA • Unsurpassed transcription through difficult secondary structures at elevated temperatures (up to 65°C) • Efficient generation of full-length cDNA, up to 14.1 kb |
| Packaging | Provided with a vial of 10× reaction buffer. |
| Legal Information | Purchase of this product is accompanied by a limited license for use in the Polymerase Chain Reaction (PCR) process for research purposes only and in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by an up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., and authorized thermal cycler. |
| Unit Definition | One unit incorporates one nanomole of TMP into TCA precipitable material in 10 min using polyadenylic acid as template and oligo(dT)12-18 as a primer. |
| eAMV is a trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. |
Properties
| concentration | 20 units/μL |
| shipped in | wet ice |
| storage temp. | −20°C |
Safety
| WGK Germany | 3 |
Related Products
| Related product | R5277, RT-PCR Protocols |
References
| reference | Brooks, E.M., et al., Secondary structure in the 3′-UTR of EGF and the choice of reverse transcriptases affect the detection of message diversity by RT-PCR,. Biotechniques 19, 806-812, (1995) Abstract |
| Tosh, C., et al., One-tube and one-buffer system of RT-PCR amplification of 1D gene of foot-and-mouth disease virus field isolates. Acta Virol. 41, 153-155, (1997) Abstract | |
| Dukas, K.P., et al., Quantitation of changes in the expression of multiple genes by simultaneous polymerase chain reaction,. Anal. Biochem. 215, 66-72, (1993) Abstract |
