MCDB 423 Lecture Notes - Lecture 2: Methyltransferase, Ampicillin, Methyl Group
9 Oct 2020
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Extracellular cysteine labeled by b" will be reduced. Targeted protein is then immunopreciptated and the amount of incorporated biotin (b) label assessed on blots. The location of the introduced cysteine residue can be deduced by comparison of the amount of biotin label incorporated with or without the pre-labeling protocol. Ampli cation occurs exponentially: at each step of pcr there are 3 types of dna present, what are the three different types of pcr dna segments, segments that are in the process of extension (unknown. Full length template length: segments of xed length equal to distance between primers. Properties of pcr enzymes: pfu compared to taq, pfu has proof reading and higher delity (less errors) Taq cheaper and faster, but not as good at amplifying sequences that are longer, and adds primer to 3" which can lead to frame shift mutation: pfu is less likely to introduce mutation (we would use pfu) We would use: quickchange ii site directed mutagenesis.
