Dna Purification Lab Report

To purify the PCR amplicon, both digested wt-goi and mut-goi from other components. A purified digested gene product is more advantageous for the DNA ligation, in which the gene products with sticky ends will be inserted to a plasmid vector. Also, to transform E.coli DH5α cells by introducing the plasmids DNA which contains the gene of interests into the E.coli strain(DH5α). The plasmid DNA can replicate inside the transformed E.coli DH5α cells, only successful transformed cells can produce the protein that is resistance to kanamycin, this allows for the selection of successful transformed cells.

2. Overview of experiments
DNA purification
Maintain the PCR volume of 50µL-100 µL. Pipet the elution buffer in the center of the PureLink®
Spin Column (PSC) and perform an incubation. Add B2 to PCR reaction. Add this sample to the PSC. Centrifuge the PSC. Replace the PSC into the wash tube. Add W1 with ethanol to the PSC. Centrifuge the PSC to remove any residual W1. Place the PSC in an elution tube. Add E1 or distilled water to the PSC. Incubate and centrifuge the PSC. The recovered elution volume should be around 48 µL. Keep the DNA sample in ice. After this step, a purify DNA sample should be obtained.
There are three tubes, each include the wild type gene of interest, the mutant goi, and one without the insert DNA to act as