March

Date	Member	Experiment	Result
2022/3/19	L.X.R.	(1) Transfer MP6 to S2060; (2) Measure the concentrarion of phage.	(1) No single colonies existed for the concentration of plasmids was a little bit high; (2) Phage plaques were not obvious.
2022/3/20	C.Q.Y.	Assemble the PACE units; validate the operation of the units.
2022/3/23	C.Q.Y.	Make 10 L of solution A and 1 L of solution B of DRM; autoclave the solution A; sterilize the solution B by using bacterial filter membranes and syringes; autoclave all instruments used to construct the PACE units.
2022/3/24	C.Q.Y.	Autoclave and dispense 1 L of DRM liquid media.
2022/3/26	C.Q.Y.	Pick the single colonies cultured by G.P.Z. and 8 gradient dilution cultures were performed in the DRM media with required antibiotics according to the protocol.
2022/3/27	L.X.R.	(1) PCR: M13 backbone, artificial sytheic section T7RNAP; (2) Electrophoresis and gel extraction; (3) Transfer MP6 into S2060.	The results of gel extraction were normal; the results of transformation were not ideal for the sizes of colonies were not the same.
2022/3/27	C.Q.Y.	(1) Measure OD600 of gradient dilution bacterial solution by microplate reader. Take proper concentration of bacterial solution to the conical flask and culture it for 2.5 h and transfer it to chemostat; (2) Make 10 L of DRM liquid media with antibiotics by using solution A and solution B. Construct the units. Run the chemostat in the light incubator at the first floor; (3) Dealing with a number of abnormal unit operations.	Time and results of OD600: 0.595(22:09), 0.284(23:09).
2022/3/28	C.Q.Y.	Take a number of samples from the chemostat to measure the OD600; adjust the setting of the units	Time and results of OD600: 0.139(11:09), 0.195(16:00), 0.925(21:05).
2022/3/29	L.J.J.	Transfer MP into the bacteria. Paint the bacterial solution onto the plates and put them in the biological shaker at 220 rpm, 37℃ for 12 h.	No colonies grew on the plates til 9:00 p.m.
	Z.Y.H., N.X.H., T.W.H.	PCR	Wrong section.
	L.X.R.	Transfer MP6 into S2060 (3 repetitions).	The bacterial colonies appeared after 24 h. Colony morphology was normal, but it was contaminated with other bacterial.
	C.Q.Y.	(1) Anomalous floating of tubes in a constant calorifier substrate bottle. Readjust the units; (2) Set the input speed of chemostat at 1 unit volume per hour. Set the light incubator at 40℃； (3) Take samples from chemostat to measure the OD600.	Time and results of OD600: 0.500(13:46), 0.636(14:39), 0.529(15:35), 0.680(16:47), 0.611(17:57); the constantiser is basically considered to be in stable operation.
2022/3/30	Z.Y.H., L.J.	PCR	Wrong section.
2022/3/30	L.X.R.	(1) Homologous recombination of the purified fragment in March, 27th. Operate the phage infection experiment; (2) Take the S2060 bacterial solution into the tubes and put them in the biological shaker. It was prepared to estimate the mutation ratio of MP6; (3) Make the arabinose solution and glucose solution and were sent to be sterilised in a high temperature.
2022/3/31	L.X.R.	(1) Dilute the bacterial solution 1000 times. Take the bacterial solution into basic media or 2xYT media respectively until the OD600 became 0.5~0.7. Divide the samples into two conical flasks and take 25 mL of bacterial solution into every flask; (2) 100 mM D-Glucose was added into one of the flasks. The same quantity of 100 mM L-Arabinose was added into another flask. Put them in the biological shaker at 37℃ for 24 h.	Put the bacterial solution in the biological shaker for 7 h.
2022/3/31	C.Q.Y.	Paint the plates with new SP made by G.P.Z.; operate the PCR for the phage; culture the bacteria in the biological shaker; electrophoresis.	The results did not match the theory.