AMV Reverse Transcriptase (Promega) .

AMV reverse transcriptase 5× reaction buffer (M515A) : The 5× reaction buffer provided with AMV reverse transcriptase is composed of 250mM Tris-HCl(pH8.3,25 ℃),250 mm KCl,50 mm MgCl2,2.5mM spermidine and 50 mm DTT.

Enzyme storage Solution: AMV Reverse Transcriptase (AMV-RT)

The storage solution consists of 200mM potassium phosphate (pH7.2, 4 ℃),0.2% Triton X?-100,2mM DTT, and 50% glycerol.

Source: Enzyme purified from avian myeloblastoma virus particles.

Storage condition: Store at -20℃. Avoid repeated freezing and thawing, and avoid exposure to environments with frequent temperature changes. Please check the expiration date on the product label.

Activity unit definition: The amount of enzyme required to catalyze the conversion of 1nmol deoxynucleotide into an acidic precipitable substance within 10 minutes at 37 ° C is one activity unit. The reaction conditions were: 50mM Tris-HCl(pH8.3),40mM KCl,8.75mM MgCl2, 10mM DTT,0.1mg/ml acetic acid BSA,1mM radiolabelated dTTP and 0.25mM poly(A):oligo(dT). Please check the activity unit concentration on the product label.

Attention:

1. AMV reverse transcriptase 5×buffer is only used for the synthesis of the First Strand of cDNA. It does not contain deoxynucleotides, so it cannot replace Promega RiboClone AMV RT first-strand 5×buffer(Part # C121A). C121A is a component of the Universal RiboClone cDNA System (catalog number C4360) containing four DNTPS. Promega's Access RT-PCR System(catalog number A1250) is a kit that uses AMV reverse transcriptase and Tfl DNA polymerase to integrate reverse transcriptase and PCR in one step without the need for an intermediate operation. The buffer is different from the AMV reverse transcriptase 5× reaction buffer. So these two buffers are not interchangeable.

2. AMV reverse transcriptase 5× buffer cannot be used for M-MLV reverse transcriptase.

3. If you are going to use Taq DNA Polymerase for PCR reaction, then you can directly add this reverse transcription reaction mixture in volume less than 10ul. If you are using GoTaq? DNA Polymerase (catalog number M3001) or PCR Master Mix (catalog number M7501) then you can directly add this reverse transcription reaction mixture up to 25μL into a 50μL PCR system.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.