Revision as of 17:56, 7 August 2014

Thursday 7th August

In progress

'by Melanie'

we use the same protocol as yesterday but we have done 8 tubes to apply a gradient in the thermocycleur during the third step:

PCR protocol: 98° --> 2min

5 PCR cycle:


time	10sec	20sec	45sec
temperature	98°	(depending on the tube - see the gradient)	72°

25 PCR cycle


time	10sec	20sec	45sec
temperature	98°	72°	72°

and last step: 72° during 10min

but we don't have any results

'by melanie' Transformation of competent cells 5mg1655) by electroporation (pJBEI6409) [1] We do some stock

by Juliette & Terry

From liquid culture made the EDIT !!!!

by Sean & Pierre

for this PCR, five tubes of each of the following Bio-bricks were prepared.

BBa_K517003

BBa_K762100

Tubes were placed in PCR machine with the following parameters.


Cycle step	Temperature	Time	Cycle
Initial denaturation	98°C	1 min	1
Denaturation	98°C	15 s	25 - 30
Annealing	52°C	25 s	25 - 30
Extension	72°C	45 s	25-30
Final extension	72°C	10 min	1
Final extension	8°C	hold	1

Members present:

Instructors and advisors: Alice.
Students: Eugene, Fabio, Hoang Vu, Juliette, Melanie, Pierre, Romain, Sean and Terry.

@@ Line 66: / Line 66: @@
 [https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_cells_by_electroporation]
 We do some stock
-==== PCR of BBa_K517003 and BBa_K762100====
-''by Pierre & Sean''
 === Extraction of the Genomic DNA===