In this issue:

(1) CyClone – universal ligase-free cloning
(2) Quantitative PCR: quite some nice new gimmicks

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CyClone – the new time-saving ligase-free cloning kit.
Eliminate all the shortcomings of conventional cloning by using the new CyClone ligase-free cloning system!

Together with optimized polymerase and buffer, CyClone offers significant advantages compared to conventional cloning or other ligase-free cloning methods:

• Compatible with any vector that can be linearized – use your standard cloning vector without any need of specialized plasmids/sequences


• High cloning efficiency with 95 - 99 % of positive clones


• Very efficient blunt end cloning and directed cloning into single-cut vectors


• Fast and convenient vector and insert preparation


• Any enzyme can be used for vector linearization, even when its restriction site is present in the insert


• CyClone does not add additional sequences to vector or insert

With CyClone cloning consists of only two simple steps:

• 1^st: Creation of a PCR product with ends homologous to the cloning vector
• 2^nd: Annealing and fusion of insert and vector in a circularization reaction (Fig. 1)

Figure 1: The CyClone cloning procedure: A PCR fragment is fused to a vector with homologous ends, yielding a plasmid useable for transformation in just one step.

• Ready-to-use formulations for EvaGreen™- and probe-based qPCR


• NEW: Blue dyed versions of all mixes to facilitate pipetting into white plasticware


• GreenMaster mixes optimized for high-resolution DNA melting curve analysis (HRM)


• Highest sensitivity for detection of ultra-traces of template DNA


• Broad dynamic range of up to 6 orders of magnitude


• Compatible with all common block-based real-time PCR cyclers

Mastermixes with EvaGreen™ Long-term stable and optimized for HRM		Mastermixes for probe-based qPCR Highly specific and optimized for multiplex applications
qPCR GreenMaster with UNG		qPCR ProbesMaster with UNG
qPCR GreenMaster		qPCR ProbesMaster
qPCR GreenMaster with UNG/ROX		qPCR ProbesMaster with UNG/ROX
qPCR GreenMaster with ROX		qPCR ProbesMaster with ROX