Enhanced Avian Reverse Transcriptase [eAMV™ RT]
SIGMA/A4464 - For reverse transcription at higher temperatures & rare mRNAs
MDL Number: MFCD01634820
Product Type: Chemical
eAMV™ RT efficiently reverse transcribes mRNA into cDNA up to 14.1 kb in size.
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 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.
Lanes
1. Marker
2. 908 bp PCR product, 2,180 transcript size
3. 796 bp PCR product, 5,760 transcript size
4. 708 bp PCR product, 6,875 transcript size
5. 608 bp PCR product, 9,970 transcript size
6. 499 bp PCR product, 10,980 transcript size
7. 396 bp PCR product, 12,890 transcript size
8. 300 bp PCR product, 14,150 transcript size
9. Negative Control (No RT)
10. 349 bp PCR product, 1,060 transcript size
11. 452 bp PCR product, 880 transcript size
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 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.
Lanes
1. Marker
2. 908 bp PCR product, 2,180 transcript size
3. 796 bp PCR product, 5,760 transcript size
4. 708 bp PCR product, 6,875 transcript size
5. 608 bp PCR product, 9,970 transcript size
6. 499 bp PCR product, 10,980 transcript size
7. 396 bp PCR product, 12,890 transcript size
8. 300 bp PCR product, 14,150 transcript size
9. Negative Control (No RT)
10. 349 bp PCR product, 1,060 transcript size
11. 452 bp PCR product, 880 transcript size
| color | colorless |
| concentration | 20 units/μL |
| feature | dNTPs included: no |
| hotstart: no | |
| form | crystalline powder |
| input | purified RNA |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| technique(s) | RT-PCR: suitable |
| usage | sufficient for 50 reactions |
| Application: | Enhanced Avian Reverse Transcriptase (eAMV RT) is used to transcribe RNA into DNA, and facilitates efficient mRNA template driven sysnthesis of cDNAs. This is due to the abillity of this enhanced AMV-RT to transcribe large mRNA templates, to transcribe through difficult secondary structures, and to detect low abundance mRNAs by RT-PCR. |
| Application: | Enhanced Avian Reverse Transcriptase [eAMV™ RT] has been used for reverse transcription of total RNA to synthesize cDNA during quantitative reverse transcription PCR (RT-qPCR) analysis. |
| Biochem/physiol Actions: | Reverse transcriptase catalyzes RNA template incorporation of dNTPs into complimantary DNA through phosphodiester bond formation. |
| 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 • Produces first strand cDNA ready for PCR amplification |
| General description: | eAMV™ Reverse Transcriptase is an enhanced form of Avian Myeloblastosis Virus (AMV) RT that synthesizes a DNA strand complementary to RNA, DNA, or an RNA:DNA hybrid. This exceptionally robust AMV RT has greater thermostability than standard AMV or Moloney murine leukemia virus(M-MLV) reverse transcriptase. eAMV™ RT is an ideal enzyme for producing high-quality full-length cDNA from total RNA or poly(A)+ RNA and is also efficient at transcribing long targets. |
| Legal Information: | eAMV is a trademark of Sigma-Aldrich Co. LLC |
| 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. |
| Packaging: | Provided with a vial of 10× reaction buffer. |
| 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |