Synthetic biology build and cloning (Tom) Flashcards
What is synthetic biology?
engineering of biology to make novel proteins/ systems/ organisms
why was synthetic chemistry important ?
the creation of urea synthetically, essentially birthed modern chemistry
give 2 examples of previous genetic engineering
- selective breeding
-cut and paste: PCR/ligase/ RE’s
outline creation in syntehtic biology
-Design: solution to problem
-Build: model/ parts
-Test: assays/analytics
-Learn
what is reverse engineering?
start with the system and take it apart to see the origin. move backwards.
What is forward engineering?
deisgn base don problme. build and model and predict.
whats abstraction
simplify and build model removed unnecessary infor/ detail
whats modularity
Modular parts that can be swapped out etc.
what is characterisation
Knowing how and why DNA works and context it worse in.
whats standardisation
standard parts/ methods/ proceedures/sharing/ results.
what are biobricks?
prefix and suffix flank every region, contain RE sites.
enzymes cut here
anneal and leave a scar
scar sequence edited out
bio brick +/-‘ves
+ easy
+standardised
- cant predict were scars form
- no diversity have to restart form beginning for new.
What is golden gate cloning?
uses type IIS RE’s to specifcally cut
DNA ligase rejoin
careless
golden gate +/-ves
+scarless
+quick
-need careful planning in advance
What is Gibson assmebly?
2 complementary fragments join together
PCR amplify fragments
Exonuclease digest 5’ end leaving 3’ overhangs
DNA polymerase fills gaps on overhnags using ssDNA and dsDNA as a primer
ligase seal nicks.
