What are the causes of vector and insert appearing smeared in a gel electrophoresis for double enzyme digestion?
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The causes of vector and insert appearing smeared in a gel electrophoresis for double enzyme digestion can be attributed to several factors. One possible cause is incomplete vector double digest, which can lead to difficulties in identifying the correct linear fragment in agarose gels . Another cause could be the size of the insert DNA, as the complexity of deciphering the fragment pattern increases with larger insert sizes . Additionally, the trial and error method used to determine the ordered arrangement of fragments in each digest can introduce uncertainty, especially when there are adjacent restriction fragments punctuated by the same restriction site . Gel electrophoresis itself can also contribute to smearing, as mechanical damage to DNA fragments can occur during purification with conventional affinity-based gel extraction methods .
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01 Jan 1995 | The paper does not provide information about the causes of vector and insert appearing smeared in a gel electrophoresis for double enzyme digestion. |
| The provided paper does not mention the causes of vector and insert appearing smeared in a gel electrophoresis for double enzyme digestion. |
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