RTL reverse transcriptase

RTL reverse transcriptase (RNaseH+)is an RNA-directed DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. RTL reverse transcriptase contains intact RNase H activity and lacks 3´ → 5´ exonuclease activity, RTL reverse transcriptase is particularly well suitable for use in RT-LAMP and first strand cDNA synthesis.

Storage and transportation: Store at -20 ℃ and transportation＜0℃

Unit definition: One unit incorporates 1 nmol of dTTP into acid-precipitable material in 20 minutes at 50°C using poly(A)•oligo(dT)25 as template-primer.

Quality Control Assays

• Endonuclease Activity: Incubation of 30 U of enzyme with 4 μg pUC19 DNA for 4 hours at 37°C resulted in no detectable degradation of the DNA as determined by gel electrophoresis.

• Exonuclease Activity: Incubation of a 50 µl reaction containing a minimum of 30 U of enzyme with 1µg Lambda-HindIII DNA for 16 hours at 37℃ yields no detectable degradation.

• RNase Activity: Incubation of a 10 µl reaction containing 30 U of enzyme with 1μg of MS2 RNA transcripts for 1 hours at 37°C resulted in no detectable degradation of the RNA as determined by gel electrophoresis.

Reaction setup:

cDNA Synthesis Protocol

a:

Thermocycling Conditions for a routine reaction：

ⓐ If Random Primer Mix is used ， an incubation step at 25℃ before 42 ℃.

ⓑIf target primer mix is used, an incubation step at 42℃-55℃ for 10 min-30 min.

RT-LAMP Protocol:

Vortex and centrifuge briefly to mix. Constant temperature incubation at 60°C-65°C for 1 hour.

Precaution

1) RTL/BstL Buffer can be dissolved at room temperature. After dissolution, it should be stored in an ice box or ice bath. After use, it should be stored at -20℃ immediately.

2) For your safety and health, please wear a lab coat and disposable gloves.