User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/05/05

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

{| width="800"
 * style="background-color: #cdde95;" align="center"|
 * style="background-color: #cdde95;" align="center"|




 * align="center" style="background-color: #e5edc8;" |

title=Search this Project


 * colspan="2" style="background-color: #F2F2F2;" align="right"|Customize your entry pages 
 * colspan="2"|
 * colspan="2"|
 * colspan="2"|

May 05th, 2010
Today I made a PCR of the fragment of 100bp obtained from the restriction with EcoRI and PstI of plasmid pSB1AK3. I prepared 4 different samples each obtained from a different colony.

This PCR was made following the same protocol from the last time with a few changes: I put 1μL of template DNA without any dilution and 17.5μL of MgCl2 (to avoid the quenching of all the Mg+ by the excess of DNA).

I just used a positive control; a PCR mix that only lack Taq Polymerase.

When the PCR was accomplished I made an agarose gel of 1.5% to prove my results.

At least the PCR was successful; I obtained 4 lines of 100bp corresponding to the fragment containing the double terminator. We get a double terminator (BBa_B0015).

However we also obtained some smaller bands and we don't know what are them; they could be due to inespecific primer annealing.


 * }