New answers tagged cloning
I assume that we are talking about eukaryotic genomes which are split into introns and exons. Depending on what you want to do with your cloned gene, you need different strategies. The method is based on the assumption that the gene sequence has only very little differences to the known sequences of other organisms. If you want to express your gene from a ...
To know if the ends are compatible see compatible cohesive ends (isoschizomers) from NEB website. If you are unsure then do a blunt end ligation. For blunting a sticky end (From the comments): Use Klenow or Pfu polymerase (or any proofreading/high fidelity polymerase). You can use Taq if you intend to to a TA cloning.
Cloning of animals typically involves: Taking nucleus from healthy adult cell of animal to clone Taking a healthy egg and replace the egg's nucleus with nucleus from source animal. Implanting egg in female, stimulating the pregnancy process. This same process should work on humans BUT: "Is it really possible to create humans in lab?" No. Animal cloning ...
E.coli strains that have dam or dcm methylases can methylate plasmid at adenines or cytosines respectively. See here — It is a NEB web page but has links to the cited references. DH5α has both the methylases- dam+ dcm+ , BL21 is dam+ dcm− and ET12567 is dam− dcm−
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