Why is a single tube not four tubes used in dyelabeledtermin

Why is a single tube (not four tubes) used in dye-labeled-terminator sequencing reactions? In comparison to the wild type GFP (GFPwt), what nucleotides and amino acids have been changed in GFPuv?

Solution

1. A single tube is used in dye labeled terminator sequencing because the seqeuncing will tell the seqeunce of four nucleotide bases present in the particular DNA in a single reaction by using dye tagged ddNTPs (dideoxynucleotide triphosphates). Each of the base is labbeled with different color in ddNTP and this ddNTP when incorporate in the PCR amplification reaction the reaction will stop at that point and procude the color based on which ddNTP is resposible for chain termination.

2.pGFPuv is a variant of GFPwt that has been optimized for brighter fluorescence when excited by standard UV light. And this is achieved by triple mutations of Phe99Ser/Met153Thr/ Val163Ala in the \'\'cycle 3\'\' variant of GFPwt.