Team:UNICAMP-Brazil/Notebooks/September 14

ColiGuard

Culture growth of bacteria containing the F plasmid

• From a permanent culture of bacteria containig the F plasmid, a sample was innoculated into liquid LB-AMP medium and let grown O/N at 37°C.

Gabriel

finO and finP's transformed bacteria

• All of yesterday's transformed and plated cells grew in the media!

• We then selected 10 colonies of each transformation (finO+pSB1A3 and finP+pSB1A3) in order to confirm them. We inoculated those colonies separately into liquid LB-AMP medium, and let then grow O/N at 37ºC.

• We will then proceed to the minipreps of the cultures and, once we could make another PCR for the inserted fragments, we will be able to confirm that both finO and finP are indeed in biobrick format.

Marcelo

PY Promoter - Purification of the digestion reactions

• We purified the digestion reactions of PY1 and PY2 with the Invitrogen's PureLink™ PCR Purification Kit, following the manufacturer's protocol without modifications.

• <p style=”text-align:justify;”>We also prepared an agarose gel to run the digestion reaction of BBa_J23100. The band in the gel was with the expected size. However, it is not possible to confirm the digestion by the size in the gel because the difference between the plasmid BBa_J23100 with and without the fragment excised by XbaI and SpeI is too small.

• <p style=”text-align:justify;”>We excised the band from the gel and used the Invitrogen's PureLink™ Quick Gel Extraction Kit to perform the purification, following the manufacturer's protocol without modifications (Protocol 7).

Fabiana and Leonardo