5x Sequencing Buffer

Enzovera 5× Sequencing Buffer (EV-SGR-003) is an optimized 5× reaction buffer formulated for use with Enzovera Sanger sequencing terminator mixes. It stabilizes cycle sequencing reactions, improves peak height uniformity, and reduces background signal across the full read range. The buffer is compatible with all BigDye-format sequencing protocols and is used at 1× final concentration in the sequencing reaction.

Function in cycle sequencing. The buffer provides the optimal ionic environment for the cycle sequencing polymerase, balancing magnesium availability, pH stability across the 96°C denaturation / 60°C extension thermal cycle, and the chemical environment required for efficient ddNTP incorporation. Diluting the dye terminator mix with separate sequencing buffer also allows users to economize on terminator chemistry — reducing the cost per reaction by extending the active terminator volume while preserving signal quality and read length.

Half-reaction and quarter-reaction protocols. For high-throughput laboratories looking to reduce sequencing cost without sacrificing read length, the buffer enables reliable half-reactions (2 μL terminator + 2 μL buffer) and quarter-reactions (1 μL terminator + 3 μL buffer) on most templates. Read lengths of 700–900 bases are routinely achievable with half-reactions, and 600–800 bases with quarter-reactions, depending on template quality. Full-reaction (8 μL terminator) is recommended for difficult templates or maximum read length.

Compatibility. The 5× Sequencing Buffer is compatible with VeraDye™ Terminator v3.1 Cycle Sequencing Kit (EV-SGR-002), with predecessor Enzovera terminator chemistries, and with third-party BigDye-format cycle sequencing reagents. The buffer formulation is non-proprietary in its chemical components and is intended as a universal cycle sequencing diluent.

Recommended applications:

• Dilution of cycle sequencing terminator mixes for half- and quarter-reaction protocols
• High-throughput sequencing workflows requiring per-reaction cost reduction
• Buffer replenishment for in-house cycle sequencing master mix preparation
• Standardization of reaction conditions across multiple terminator lots

Pack sizes: 5 mL and 30 mL. Store at −20°C. Stable for 24 months from date of manufacture in single-use aliquots.