What are the PCR requirements for In-Fusion reaction?
What are the PCR requirements for In-Fusion reaction?
PCR-amplified DNA must be purified prior to the In?Fusion Cloning reaction. This can be accomplished with one of the following options:
1. NucleoSpin Gel and PCR Clean-Up
-Gel extraction enables selection of specific DNA fragments of the desired size from background PCR byproducts or other contaminants.
-Column purification is appropriate if PCR did not produce a background smear.
2. Cloning Enhancer (CE)
-This proprietary enzyme mix removes background plasmid DNA and PCR residue.
-CE is appropriate for PCR that results in a single fragment of the expected size, without a background smear.
-CE is a convenient tool for high-throughput (HTP) applications that employ highly optimized PCR cycling conditions and primers that generate clean DNA fragments of the expected size.
Note: In most cases, CE treatment does not require additional column purification or gel extraction. However, to ensure better cloning results, PCR-linearized vectors may require a combination of CE treatment followed by gel extraction to separate a linearized vector from possible PCR byproducts.