Purification of pTet and double terminator biobricks
Once we digested those biobricks yesterday, we could proceed today to the purification of the target band from an agarose gel.
We used Invitrogen's Purelink Quick Gel Extraction Kit, following the manufacturer's protocol (Protocol 7), without modifications.
Marcelo
PY Promoter - Digestion and ligation reactions
Today we digested PY1 + pGEM with EcoRI and SpeI to excise PY fragment.
We also digested the plasmid BBa_J23100 (that one with the RFP reporter) with EcoRI and SpeI. With this plasmid digested with this 2 enzymes it is going to be possible to insert our digested PY fragment into this plasmid.
This digestion reactions lasted for 3 hours.
After the digestion we performed the ligation reaction of PY1 + BBa_J23100 following Protocol 11.
Fabi and Léo
YeastGuard
Dephosphorylation - CIAP test
The E.coli didn´t grow in any of the plates again. =[ We discovered that the electrocompetent E.coli that we used yesterday were dead! So we transformed another electrocompetent E.coli using the rest of the ligation reactions we had. We extremely hope to find colonies tomorrow!
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