Additionally, Creon may be feeling quite defensive since he is in the wrong: the brothers fight and kill each other because Creon supports Eteocles' refusal to honor a legal, joint royal power-sharing arrangement with twin brother Polyneices.
Thermostable DNA polymerases that have a proofreading function, such as Pfu and Tli (a) Â® -T Vectors if they are first tailed with dATP using Taq DNA Polymerase (1). This A-tailing protocol can also be used to subclone DNA fragments generated by restriction enzymes which produce blunt ends (1).igure 2. Analysis of recombinants obtained after cloning of an A-tailed PCR fragment into the pGEM Â® -T Easy Vector. A 1,200bp fragment was generated by amplification with Pfu DNA polymerase. This fragment was tailed using the alternative procedure described in Figure 1B.
Aliquot 22.75 Âµl of the PCR Master Mix without Taq Polymerase to the No EnzymeControls.10. Add Taq Polymerase (0.25 Âµl x number of remaining reactions) to the PCR âMasterMixâ and mix. Aliquot 23Âµl of the mixture into tubes containing experimental samplesand Control Template dilutions.