User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/06/22

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PCR success!!!


  • I ran a gel in the morning to check that my PCR product was there. Fortunately it was, as is shown in the gel:




  • The lanes in the gel are as follows:


1. Ladder.
2. Positive control.
3. Negative control.
4. PCR 1 using Prefix and the primer for the mutation (Reverse) with diluted DNA.
5. PCR 1 using Prefix and the primer for the mutation (Reverse).
6. PCR 2 using primer for the mutation (Forward) and Suffix with diluted DNA.
7. PCR 2 using primer for the mutation (Forward) and Suffix.
8. Purified plasmid J23106.


  • As the product was there and the punctual mutation technique needs a lot of DNA, I decided to repeat the PCR realized yesterday. This time I only did the PCR1 and PCR2 (3 tubes each).


  • To check that everything went well, I ran another gel.




  • This time the lanes were:


1. Ladder.
2. Positive control.
3. Negative control.
4. PCR 1.
5. PCR 1.
6. PCR 1.
7. PCR 2.
8. PCR 2.


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