Part $4$: RNAi experiment $(25$ points$)$

$25.$ As you are making the construct, you realize that you need to make a batch of RNAi plates for your experiment. The recipe for making MYOB plates is found below. You will make $200$ mLs of MYOB broth instead of $1$ L $.$

You have the following reagents in the lab. How will you use those reagents to make $200$ mLs of solution? $(6$ points$)$

RNAi plates $($MYOB $+$ IPTG and Carbenicillin$)($for maintenance of worm strains$)$:

Per $1$ L :

$2\mathrm{.}0$ g NaCl

$0\mathrm{.}55$ g TrisHCl

$0\mathrm{.}24$ g TrisOH

$4\mathrm{.}6$ g Bactotryptone

$20$ g agar

$1$ ml cholesterol $(8m\frac{g}{m}l$ in $95\%$ ethanol$)$

$1L{H}_{2}O$

Autoclave without the cholesterol, IPTG and antibiotics and then add cholesterol, IPTG to a final concentration of $1$ mM and carbenicillin $($similar to ampicillin$)$ to a final concentration of $50g$ per mL $.$

You have powder NaCl $,$ TrisHCl, TrisOH, Bactotryptone, agar and cholesterol, carbenicillin and IPTG.

It is easiest to add a solution of IPTG, carbenicillin, and cholesterol after autoclaving and often people prepare a $1000$X stock solution of each of these reagents. Please describe the steps you would take to prepare the plates including the stock solutions.You have now made MYOB plates seeded with the bacteria that has your siRNA construct. Describe the steps that you will do to knockdown pop$-1$ in wild$-$type $($N$2)$ worms.

$26.$ In setting up your RNAi experiment, what should you use for controls and what does each of the controls tell you? $(4$ points$)$