840:153g:Projects/project13/2010/11/18

Tuesday

• We reran the gel that we ran on thursday of last week. This time we cut our promoter part BBa_J23102 with just the spel enzyme. This would tell us if there is a binding site in the middle by the size of band that was released. If there was a 600 bp band then there would have been a xbal or spel binding sight in the backbone, if there was nothing then it was a scar. However we did not see a band. Thus there is not binding sight in the middle of the promoter.

• We also ran a gel on our ligated part again. We cut one set with ecori and spel, and another set with xbal and spel. We also included some undigested DNA for a comparision. We also had to change the amount of DNA in our wells because we were running low. So instead of having 12μL like last week, we only had 7μL. However, We saw little differnace between the two differnt digests and the undigested DNA. We looked back and saw that the orginal part BBa_J23119 was working correctly but for some reason it is unable to show the 50bp band that it did before.

• So from left to right we have the low range ladder then colonies 4,11,12,and 13 cut with xbal and spel. In the 4 wells we have the undigested colonies 4,11,12, and 13. The next four wells were cut with ecori and spel and are from colonies 4,11,12,and 13. Then their are two ladders Low range, and then 100bp ladders.
• next is our part BBa_J23102 cut with just spel and then the undigest part. and then the next well is the 100bp ladder.

Thursday

• We sent BBa_K123003 (ER) and part BBa_J23119 to be sequenced to see if these are the right parts and to figure out what step possibly went wrong.