From OpenWetWare
Jump to navigationJump to search
Our goal is to upregulate production of dsRNA of our worm gene of interest in pL4440 vector plasmids containing that gene in HT115(DE3) bacteria. <br><bR>
'''To induce your cultures, the lab specialist or your instructor:''' <br>#Add Added 5 μL of 0.5 M IPTG to your culture. What is the effective concentration of IPTG in your culture?
#Put your culture back in the 37°C incubator in the spinning wheel for approximately 3-4 hours.
#After the lab introduction, While we will head to have our journal article discussion in a more comfortable room to discuss , the papers you were assignedbacteria will be working hard expressing T7 RNApolymerase and transcribing complementary mRNA of ''C. elegans' bli-1".
'''To do after induction is complete:'''
'''4 days before next lab:'''
#Come into lab and find your stack of plates. For Friday, Monday and Tuesday sections your plates will be at room temperature by your worm picks. For Wednesday and Thursday sections your plates will be in the white refrigerator in the lab - please The first person to come in on Saturday or Sunday should remove the whole worm plate box and leave it on the counter near the worm picks so that the plates will be pre-warmed for your classmates. You should warm the your own plates up to room temp before transferring your worms (hold them in your hands or put them in the 37C incubator by the door for a few minutes ) or you will shock the worms with the coldmedium.
#On 2 of the experimental plates add 2 L4 wild type (N2) hermaphrodites
#On 2 of the experimental plates add 2 L4 ''rrf-3'' hermaphrodites


Navigation menu