Approved: Fortect
In some cases, your computer may display a cloning troubleshooting message. There can be several reasons for this problem.
| PROBLEM | REASON | SOLUTION |
|---|---|---|
| Several or no transformants | cells are not viable | Transform large uncut plasmid (eg pUC19 … |
| Little or no transformants | Bad antibiotic, antibiotic … | Confirm antibiotic and antibiotic … |
| Few transformants or they are not present | The DNA fragment of interest may be toxic … | |
| Several transformants or none | |
Follow manufacturer’s specifications … |
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Incomplete digestion of the PCR product or its absence.Incomplete digestion of the plasmid.Unexpected streaks after digestion.Too many colonies with empty vectors (ligation of the DNA fragment to the vector did not occur)No colonies or too few colonies.
Ligatures don’t work for only one of the following three reasons. First, the ends of your DNA don’t match. Second, you have a chemical inhibitor or damaged DNA (such as excessive UV exposure) that is blocking beneficial ligation. Thirdly, your vector has a high level of understanding (incomplete assimilation), and you have already ruled out this wise choice.
If two biological materials look the same, your ligature will never work. You can check the teeth whitening gel through Electrop. if vector + insert is just larger than the base vector, or digest (which just cuts the insert, not the entire vector) insertions and ncheck if you have 2 fragments.
