Sybrsafe Case Study

4. Follow-up questions

Q1. Why SYBRSafe is needed in DNA gel electrophoresis?

SYBRSafe is a dye for DNA staining to determine the presence of DNA fragment in gel electrophoresis. The SYBRSafe performs like ethidium bromide that binds to DNA fragment. SYBRSafe is excited by UV or strong light (e.g. blue light) and emits fluorescent signal which is proportional to the concentration of DNA in the gel (Thermofisher Scientific, 2011). In this experiment, SYBRSafe is used for the detection of mutant OFP gene and the corresponding Plasmid DNA, pRSET vector after restriction digestion by KpnI and HindIII. Signal strength of SYBRSafe can be one of the parameters to measure the efficiency of restriction digestion.

Q2. What is the purpose of the addition of ampicillin to the LB-agar plates?
In order words, Taq DNA Polymerase tends to develop more random mutations along DNA sequences through amplification while Platinum® Pfx DNA Polymerase tends to make a precise DNA amplification copy of DNA fragment. They can be applied in different area by means of error rate control. Taq DNA Polymerase makes more copies of DNA sequences with different mutation sites which is essential to study the effects to expression rate of a target gene in different mutation sites. Platinum® Pfx DNA Polymerase on the other hand, is able to maintain accurate amplification and ensure the specific site mutagenesis by minimizing the random error