Chinese Medicine testing
Chinese Medicine CYP
To test the influence of scutellariaon E.coli growth rate
DH5α pcDNA (ctrl)on SOCagar plates
15 ml tube、50 ml tube
96 well plate
1.Culture 4 tubes of DH5α(with pcDNA)in 15ml centrifuge tubeovernight: 2ml SOC with 2ulAMP
1.Preparesix15ml centrifugaltube, each containing 1ml of SOCand 2ul AMP
2.Put the six tubes into a 37°C water bath for five minutes.
3.After five minutes, take out the six tubes and add 1ml ofthe overnight culturedbacteria liquidinto the six tubes.
4.Culture them for another two hoursin a 37°C shakerto make sure if they cangrownormally.
5.Prepare five kinds of concentration of scutellariaon, 10μg/ml, 3μg/ml, 1μg/ml, 0.3μg/ml, 0.1μg/mlby serial dilution.
6.Prepare eighteen 50ml tubes, each contains 29.1ml of SOC,and 0.3ml schutellariaon solution(with five kinds of solution).
7.Put the eighteen 50ml tubes into the 37°C water bath for five minutes
8.Add 0.6mlbacteria liquidfrom the six tubes
9.Examine the OD of each tube every 30 minutes
From this result, we can see that the growthratesof the control and different concentration are very simular. Therefore, we know that that scutellariaonwon’t influence the growth rate of E.coli.