PhiScript – DX/DT’s incredible reverse transcriptase! It’s an engineered form of Moloney Murine Lukemia Virus reverse transcriptase (M-MuLV-RT). Point mutations are introduced in ɸScript™ reverse transcriptase so as to reduce RNAse H Activity and to improve its thermal stability. Good cDNA yields can be obtained when reverse transcribed at 37 – 60°C.Trusted by the scientific community, this enzyme is the go-to for converting RNA into complementary DNA (cDNA) with lightning speed and precision. Whether you’re exploring gene expression dynamics or diving into RNA sequencing, PhiScript has your back. Its exceptional efficiency and fidelity ensure reliable cDNA synthesis, unlocking insights into the intricate world of RNA biology.
Mutant MMLV RT for stable reverse transcription.
Reverse transcription upto 60℃ is supported.
Hig yield cDNA synthesis just under 30 minutes.
Precise transcriptions in OneStep RT-PCR reactions.
High tolerance for many PCR inhibitors/reagents.
Glycerol Free Enzymes available for Lyo Applications.
Accurate results. Simple. Incredible prices.
Ultrafast delivery across India. Less than 3 hrs for Namma Bengaluru.
Features
Source
PhiScript™ reverse transcriptase was cloned from MML Virus gene, mutated and expressed in E.Coli.
DX/DT Phiscript(ɸ) reverse transcriptase is used in cDNA Synthesis Kit and cDNA mastermix. Hence, all the data collating here with respect to both.
Total human RNA was isolated from skeletal cells. Exon junction primers were designed for human integrin mRNA transcript.
cDNA Synthesis conditions
Total RNA template (1µL=10ng), cDNA 5X mastermix(4µL each), ɸScript Reverse Transcriptase, 1µL) and make upto 20µL using Nuclease Free Water.
Cycling conditions
25℃ (5min)
42℃ (20min)– Reverse Transcription
85℃ (10min) — Inactivates RT
-20℃ — Storage
PCR Conditions
cDNA from the condition was tested and mastermix ability was checked for 3 different sizes of the same human integrin transcript ( 150bp, 650bp and 1.6kb). Appropriate forwad and reverse primers were used.
General PCR conditions:
cDNA: 2µL, For/Rev primers: 1µL each, DX/DT Extremo mastermix: 25µL and make up 50µL using nuclease free water.
PCR Cycles:
95℃ – 2min, [95℃-30s, 60℃-30s, 72℃-1min/kb]30cycles, Final extension, 72℃ for 2min.
5µL of the sample was loaded on 1.5% agarose gel and run for 30 minutes.
Reverse transcription temperature is crucial for certain application where RNA molecule is complex and contains high amounts secondary structure. cDNA synthesis with these samples are always challenging. But worry not. Phiscript (ɸ) reverse transcriptase has exceptional stability to work at higher temperatures. This is also one of the critical parameters during one step reverse transcription where non-specific amplification has to be reduced.
Total human RNA was isolated from skeletal cells. Exon junction primers were designed for human integrin mRNA transcript.
cDNA Synthesis conditions
Total RNA template (1µL=10ng), cDNA 5X mastermix(4µL each), ɸScript Reverse Transcriptase, 1µL) and make upto 20µL using Nuclease Free Water.
Cycling conditions
25℃ (5min)
37℃ (30min), 42℃ (30min), 50℃ (30min), 60℃ (15min) — Reverse Transcription (Separate vials)
85℃ (10min) — Inactivates RT
-20℃ — Storage
PCR Conditions
cDNA from the condition was tested and mastermix ability was checked for 3 different sizes of the same human integrin transcript (150bp and 1.6kb). Appropriate forwad and reverse primers were used.
General PCR conditions:
cDNA: 2µL, For/Rev primers: 1µL each, DX/DT LeoPrime mastermix: 25µL and make up 50µL using nuclease free water.
PCR Cycles:
95℃ – 2min, [95℃-30s, 60℃-30s, 72℃-1min/kb]30cycles, Final extension, 72℃ for 2min.
5µL of the sample was loaded on 1.5% agarose gel and run for 30 minutes.
DX/DT cDNA 5X performance was compared against other brands in the market. Total human RNA was isolated from skeletal cells. Exon junction primers were designed for human integrin mRNA transcript.
cDNA Synthesis conditions
Total RNA template (1µL=10ng), cDNA 5X mastermix(4µL each), ɸScript Reverse Transcriptase, 1µL) and make upto 20µL using Nuclease Free Water.
Cycling conditions
25℃ (5min)
42℃ (30min) — Reverse Transcription (Separate vials for competitors as per their instruction)
85℃ (10min) — Inactivates RT
-20℃ — Storage
PCR Conditions
cDNA from the condition was tested and mastermix ability was checked for reverse transcription of 650bp fragment. Appropriate forwad and reverse primers were used.
General PCR conditions:
cDNA: 2µL, For/Rev primers: 1µL each, DX/DT LeoPrime Mastermix: 25µL and make up 50µL using nuclease free water.
PCR Cycles:
95℃ – 2min, [95℃-30s, 60℃-30s, 72℃-1min/kb]30cycles, Final extension, 72℃ for 2min.
5µL of the sample was loaded on 1.5% agarose gel and run for 30 minutes.
Phiscript™ reverse transcriptase is powerful to yield good amount of cDNA even when initial RNA template is low or partially degraded.
Total human RNA was isolated from skeletal cells. Total RNA was diluted using nuclease free water. Exon junction primers were designed for human integrin mRNA transcript.
cDNA Synthesis conditions
Total RNA template (1µL=10ng), cDNA 5X mastermix(4µL each), ɸScript Reverse Transcriptase, 1µL) and make upto 20µL using Nuclease Free Water.
Cycling conditions
25℃ (5min)
42℃ (30min) — Reverse Transcription (Separate vials for diluted RNA)
85℃ (10min) — Inactivates RT
-20℃ — Storage
PCR Conditions
cDNA from the condition was tested and mastermix ability was checked for reverse transcription of 650bp fragment. Appropriate forwad and reverse primers were used.
General PCR conditions:
cDNA: 2µL, For/Rev primers: 1µL each, DX/DT LEO PRIME: 25µL and make up 50µL using nuclease free water.
PCR Cycles:
95℃ – 2min, [95℃-30s, 60℃-30s, 72℃-1min/kb]30cycles, Final extension, 72℃ for 2min.
5µL of the sample was loaded on 1.5% agarose gel and run for 30 minutes.
Protocol for this product can be found here.
All the products are for research purpose only. Not for human diagnostics.
