PCR, PCR Clean up, Cloning, Sub-cloning, Mutagenesis
Description of Service
PCR, PCR clean up
PCR of genomic DNA or BAC DNA are done using custom primers. Primers can also be designed at the facility. The optimization and the clean up of PCR products are done using various procedures. The cleaned up products can be directly processed for sequencing at the facility.
Transformation and Cloning
Design of cloning strategy followed by directional or non-directional clonings are performed using appropriate competent cells and vectors. Insertions sites are checked and orientations are determined by either restriction digestion and/or sequencing.
PCR products are cloned using Invitrogen’s TA cloning system.
Sub-cloning
After designing the cloning strategy, DNA is sub-cloned into appropriate expression vector followed by the preparation of the construct DNA for micro-injection into mouse zygotes.
Site-directed Mutagenesis
Mutagenesis including point mutation, insertion and deletion of single or multiple amino acids and large insertion are done using Stratagene’s Quick Change kit. This includes design and synthesis of the mutagenic primers, mutagenesis and verification by sequencing. The synthesis and the PAGE purification of the primers are outsourced to Invitrogen.