Universal cDNA Synthesis and SYBR® Green Master Mix kits

Universal cDNA Synthesis kit II

All the reagents needed for fast and convenient microRNA polyadenylation and reverse transcription in a single reaction step. By using a single universal RT reaction as template in all subsequent PCR reactions, the procedure is greatly simplified compared to systems that require microRNA-specific first-strand synthesis (Figure 1).

When used in 20µL reactions, the kit contains reagents sufficient for 32 reverse transcription reactions. Reaction volumes can be varied between 10 and 80µL (Figure 2).

Contents of the kit:

• 5x reaction buffer
• 10x enzyme mix
• nuclease-free water
• RNA spike-in template (UniSp6)

The UniSp6 RNA spike-in can be used with the Control primer set provided with the ExiLENT SYBR® Green master mix or as a positive control and/or inter-plate calibrator in Exiqon’s microRNA PCR panels.

Please note, UniSp6 is supplied in double the amount of the old Universal cDNA Synthesis kit and should be dissolved in twice the amount of water to have the same working concentration. In all other regards the two kits are identical.

ExiLENT SYBR® Green master mix

ExiLENT SYBR® Green master mix is a new high-performance PCR master mix kit specifically designed for Exiqon’s miRCURY LNA™ Universal RT microRNA PCR system. It offers improved sensitivity over the old SYBR® Green Master mix it is replacing (Product no. 203400 & 203450).

The ExiLENT SYBR® Green master mix kit is required for use with Exiqon’s new v.3 miRCURY LNA™ Universal RT microRNA PCR assays and panels, and is not compatible with older v.2 products. Results obtained using the old and new kits are comparable for delta Cq values. However, direct Cq comparisons between the two kits is not recommended. For this reason we recommend using the old kit if you are adding new samples to an existing profiling project. For new projects, the ExiLENT SYBR® Green master mix kit should be used.

The kit is available in two sizes: a 2.5 mL kit containing reagents for 500 PCR reactions (10µL reactions) and a 20mL kit sufficient for 4,000 reactions.

Contents of the kit:

• 2x ExiLENT SYBR® Green Master Mix
• nuclease-free water
• Control primer set (UniSp6)

The Control primer set is designed for amplification of the synthetic RNA spike-in provided in the Universal cDNA synthesis kit. Please refer to the miRCURY LNA™ Universal RT microRNA PCR Instruction manual for details.

Universal cDNA Synthesis kit II

The kit contains reagents for fast and reliable reverse transcription of all microRNAs into cDNA in a single reaction step. The number of pipetting steps are kept to a minimum in order to save time and minimize technical variation.

ExiLENT SYBR® Green Master Mix

This kit contains high performance reagents for quantitative real-time PCR amplification. The detection is based on SYBR® Green which allows quality control of the resulting PCR amplicon by melt curve analysis. LNA™-enhanced primer sets require specific PCR reaction conditions for optimal performance.
Exiqon's v.3 primer sets and PCR Panels have been validated using the ExiLENT SYBR® Green master mix. The use of other reagents for qPCR will affect the results. The ExiLENT master mix is NOT compatible with older v.2 products, where the old SYBR® Green master mix should be used (Product no. 203400 & 203450).

A key feature with the new ExiLENT SYBR® Green master mix is the remarkably low residual activity. This means it is possible to work reliably with Cq values up to 37 cycles, giving the platform exceptional sensitivity without the use of time consuming preamplification. Reliable profiling in the high Cq range is a key feature required for working with samples low abundant in microRNAs like biofluids (Figure 1).