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SOLVED: Suppose that you wanted to ligate X-vector ( 8 kb) with one DNA fragment ( 1 kb). The vector was digested using NheI-HF, KpnI-HF, BamHI-HF, and the DNA fragment was digested with NheI-HF, KpnI-HF, and DpnI high fidelity restriction enzymes. You used 10X NEB Buffer 4 to ligate them. You checked the ligation reaction via transformation and you got clones but not DNA fragment inserted. You have repeated the procedure 3 times but could not get the ligation to work. What are possible causes f
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