Home FAQs What are the differences between the Luna One-Step RT-qPCR products (NEB #E3006, #E3007, #M3019, #M3029 and #L4001)?

FAQ: What are the differences between the Luna One-Step RT-qPCR products (NEB #E3006, #E3007, #M3019, #M3029 and #L4001)?

These Luna products are all optimized for one-step RT-qPCR by probe detection. NEB #M3019/#M0329 are one-tube 4X master mixes The NEB #E3006/#E3007 products contain separate tubes for the 2X reaction mix and 20X enzyme mix. Also, the M3019 product includes not only dUTP, but also a thermolabile UDG, to fully enable carryover prevention. NEB #L4001 is a lyophilized version of NEB#M3019 and is available in vials or 96-well plates. NEB #E3007 and #M3029 lack a universal ROX passive reference dye. Available mixes differ with regards to format, storage temperature, and inclusion of thermolabile UDG for carryover prevention, as outlined in the table below.
 
  NEB #M3019
Luna Probe One-Step RT-qPCR 4X Mix with UDG
 NEB #M3029
Luna Probe One-Step RT-qPCR 4X Mix with UDG (No ROX)		 NEB #E3006
Luna Universal Probe One-Step RT-qPCR Kit		 NEB #E3007
Luna Probe One-Step RT-qPCR Kit (No ROX) 		NEB #L4001
LyoPrime Luna Probe One-Step RT-qPCR Mix with UDG
Format 4X Master Mix
 4X Master Mix
 2X Reaction Mix
20X Enzyme Mix		 2X Reaction Mix
20X Enzyme Mix		 Lyophilized Master Mix; rehydrate at 2-4X
Carryover prevention included Y
(dUTP and thermolabile UDG included)
 Y
{dUTP and thermolabile UDG included)		 N
(dUTP included; requires UDG addition)		 N
(dUTP included; requires UDG addition)		 Y
(dUTP and thermolabile UDG included)
Normalization dye
(universal ROX)		 Y N Y N Y
Multiplexing Y (up to 5-plex)
 Y (up to 5-plex)
 Y (up to 5-plex) Y (up to 5-plex) Y (up to 5-plex)
Maximum sample volume
(20 µl RT-qPCR reaction)		 up to 10 µl
 up to 10 µl
 up to 8 µl up to 8 µl up to 10 µl
Standard cycling condition 25°C for 30 sec. (UDG treatment);
55°C for 10 min;
95°C for 1 min;
45 cycles of 95°C for 10 sec., 60°C for 30 sec.
 25°C for 30 sec. (UDG treatment);
55°C for 10 min;
95°C for 1 min;
45 cycles of 95°C for 10 sec., 60°C for 30 sec.
 55°C for 10 min;
95°C for 1 min;
45 cycles of 95°C for 10 sec., 60°C for 30 sec.		 55°C for 10 min;
95°C for 1 min;
45 cycles of 95°C for 10 sec., 60°C for 30 sec.		 25°C for 30 sec. (UDG treatment);
55°C for 10 min;
95°C for 1 min;
45 cycles of 95°C for 10 sec., 60°C for 30 sec.
Storage temperature -20°C
 -20°C
 -20°C
 -20°C
 Room temperature (15°C to 25°C)