Home FAQs How do I choose between the first strand cDNA synthesis kits from NEB?

FAQ: How do I choose between the first strand cDNA synthesis kits from NEB?

Table 1 highlights the major advantages and applications of the first strand cDNA synthesis kits from NEB.

cDNA Synthesis Kits LunaScript® RT SuperMix Kit (NEB #E3010) LunaScript RT Master Mix Kit  (Primer-free) (NEB #E3025) ProtoScript® II First Strand cDNA Synthesis Kit (NEB #E6560) 
Components
  1. LunaScript RT SuperMix (5X)*
  2. No-RT Control Mix (5X)*
  3. Nuclease-free Water

    *Both mixes contain hexamer and d(T)10
  1. LunaScript RT Master Mix (Primer-free) (5X) 
  2. No-RT Control Mix (Primer-free) (5X) 
  3. Nuclease-free Water
 
  1. ProtoScript II Enzyme Mix (10X) 
  2. ProtoScript II Reaction Mix (2X) 
  3. Oligo d(T)23VN (50 µM)
  4. Random Primer Mix (60 µM)
  5. Nuclease-free Water
 
 
Format
 
Recommended application
Primers
Incubation

For 2-step RT-qPCR detection

  • Hexamer and d(T)10 included
    25°C 2 min; 55°C 10 min; 95°C 1 min
 
 

For 2-step RT-qPCR detection

  • Random Primer Mix
    25°C 2 min; 55°C 10 min; 95°C 1 min
For general cDNA synthesis
  • d(T)23VN or gene-specific primers
    55°C 10 min; 95°C 1 min
  • Random Primer Mix
    25°C 2 min; 55°C 10 min; 95°C 1 min

For general cDNA synthesis

  • d(T)23VN or gene-specific primers
    42°C 60 min; 80°C 5 min
  • Random Primer Mix
    25°C 5 min; 42°C 60 min; 80°C 5 min
 
Advantages ✓ Supermix
✓ Short protocol
✓ A visible tracking indicator
✓ Higher reaction temperature
✓ Higher volume of RNA input		 ✓ Flexible choice of primers
✓ Short protocol
✓ A visible tracking indicator
✓ Higher reaction temperature
✓ Higher volume of RNA input 		 ✓ Flexible choice of primers
✓ Everything included
 

 

Follow the decision tree below to choose the first strand cDNA synthesis kit based on your downstream applications.

Figure 1. LunaScript RT Master Mix (Primer-free) supports numerous applications.



By adding different primers including Random Primer Mix, d(T)23VN, and random hexamers, cDNA produced by LunaScript RT Master Mix can be ideally suited for downstream applications such as RT-qPCR detection, RT-PCR, and RNA-seq studies.