A student cloned a primer of a gene and did a recombination using a vector (yeast). The student is trying to confirm the recombination after a pcr amplification but all gels run failed to show bands. What could the student have done wrong? or not doing right???.
Hello, several reasons can cause the problem.
1.It can be problem with gene or yeast vectors itself.
2. Can be very small quantity gene in amplification or some inhibitors of amplification during PCR.
3. PCR program and conditions
4. Not correct primer.
5. Wrong pipetting
6.Not correct conditions during gel electrophoresis wrong polarity that amplification product goes wrong way or high voltage that product pass all gel during electrophoresis.
7. Visualisation problem in gel chemical or preparing problems.
8. etc.
Hello, several reasons can cause the problem.
1.It can be problem with gene or yeast vectors itself.
2. Can be very small quantity gene in amplification or some inhibitors of amplification during PCR.
3. PCR program and conditions
4. Not correct primer.
5. Wrong pipetting
6.Not correct conditions during gel electrophoresis wrong polarity that amplification product goes wrong way or high voltage that product pass all gel during electrophoresis.
7. Visualisation problem in gel chemical or preparing problems.
8. etc.
I agree with answer above, so you have to find the problem source.
Start with easy steps.
First, when you ran gels you ran a marker to see your gel is good?
Second blast your primers.
And go deep and deeper to find the problem
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