5. All of your YPD patches will be replica printed onto a MATa lawn and onto a MATalpha lawn. Each of those replicas will be incubated for 48 hours and then printed onto SD. You will therefore have two SD plates, one originating from the MATa tester replica and one originating from the MATalpha tester replica.

a. Should any of the specific patches that arise on the SD plate originating from the MATa tester replica also arise on the SD plate originating from the MATalpha tester replica? (i think the answer is yes)

b. If you have a diploid patch on your YPD patch plate, will that colony result in growth on the SD plate originating from the MATa tester replica?

c. If you have a diploid patch on your YPD patch plate, will that colony result in growth on the SD plate originating from the MATalpha tester replica?

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Jean KeelingLv2

3 Jul 2019

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In a genetic experiment:

The use and limits of RNA interference as a reverse genetics technique for knocking down gene expression were tested. The wild-type and mutants worms for the strains were selected. The RNAi feeding strains and control bacterial were used for the the RNAi experiment. The eri-1 C. elegan strains (wild-type) were used for RNAi treatment and mutant strains that were used ( unc-22 and dpy-13) C. elegans. Both carbencillin and IPTG wered added to the RNAi feeding plates and E.coli containing the RNAi transgenes were spread on the plates. The antibodic carbacillin only allows growth of bacteria which contains the RNAI construct. IPTG induces the expression of the polymerase which then transcribes the siRNA in the bacteria. 6 NGM plates without bacteria were used and 20ul of 12.5 mg/ml carbencillin concentration were spread on the 6 of the plates. The carbencillin dried and then IPTG was spread (25ug/ml) on each plate. Then 6 plates with the RNAi bacterial strains were used. Theree of plates for strain one and the other three for strain the second strain. When IPTG dried , 150 ul of the appropraite bacteria was added onto each plate. 2NGM plates (without carbenicillin or IPTG) was spread with 150 ul of OP50 bacterial culture. OP50 is the regular E. coli strain for culturing C. elegans and does not contain the RNAi constructs. These plate were used as the control plates6. Then the plates were incubated for two days. Then the adult hermaphrodites from the RNAi feeding plates and the op50 plates were removed. Then from the eri-1 wildtype stock, 2 or 3 L4 hermaphrodites or
young adults were transferred onto each RNAi feeding plate. Then L4 worms were transferred to an intermediate plate first. After they have a minute to move away from each other and the eggs, and transfered to the RNAi plates. On the two control plates seeded with OP50, 2 L4 or young adult hermaphrodites were placed there. Plates were obtain as the reference mutants (unc-22, bli-1 or unc-13). Then the plates were incubated to allow the worm to develop and lay eggs in two days. Then the adult hermaphrodites from your RNAi feeding plates and the op50 plates were removed and then placed back into the incubator.

The op500 was the negative control plate wtih 80 and 300 progeny and unc-22 had around 50. (reference plates)

Mating in the RNAi plates

Dpy (60) x wildtype (15) and unc( 70) x wildtype ( 8) progeny

the phenotype was movement and it seemed similar phenotype within the progeny.

Can you help me understand the results and what is happening genetically in the experiment ? Are the for dyp-15 x unc-22 mating cross and unc-22 x eri-1 wildtype C. elegans. Based on the data:

Briefly discuss the results of the RNAi experiment. How closely did the RNAi-treated worms phenocopy the mutant worms?

Was the gene knockdown 100% effective?

Was the RNAi treatmentequally effective for the two genes? If not, discuss this result and why that could occured.

Please be specific and detailed

Genetics HELP ASAP

Genetics HELP ASAP In a genetic experiment: The use and limits of RNA interference as a reverse genetics technique for knocking down gene expression were tested. The wild-type and mutants worms for the strains were selected. The RNAi feeding strains and control bacterial were used for the the RNAi experiment. The eri-1 C. elegan strains (wild-type) were used for RNAi treatment and mutant strains that were used ( unc-22 and dpy-13) C. elegans. Both carbencillin and IPTG wered added to the RNAi feeding plates and E.coli containing the RNAi transgenes were spread on the plates. The antibodic carbacillin only allows growth of bacteria which contains the RNAI construct. IPTG induces the expression of the polymerase which then transcribes the siRNA in the bacteria. 6 NGM plates without bacteria were used and 20ul of 12.5 mg/ml carbencillin concentration were spread on the 6 of the plates. The carbencillin dried and then IPTG was spread (25ug/ml) on each plate. Then 6 plates with the RNAi bacterial strains were used. Theree of plates for strain one and the other three for strain the second strain. When IPTG dried , 150 ul of the appropraite bacteria was added onto each plate. 2NGM plates (without carbenicillin or IPTG) was spread with 150 ul of OP50 bacterial culture. OP50 is the regular E. coli strain for culturing C. elegans and does not contain the RNAi constructs. These plate were used as the control plates6. Then the plates were incubated for two days. Then the adult hermaphrodites from the RNAi feeding plates and the op50 plates were removed. Then from the eri-1 wildtype stock, 2 or 3 L4 hermaphrodites or young adults were transferred onto each RNAi feeding plate. Then L4 worms were transferred to an intermediate plate first. After they have a minute to move away from each other and the eggs, and transfered to the RNAi plates. On the two control plates seeded with OP50, 2 L4 or young adult hermaphrodites were placed there. Plates were obtain as the reference mutants (unc-22, bli-1 or unc-13). Then the plates were incubated to allow the worm to develop and lay eggs in two days. Then the adult hermaphrodites from your RNAi feeding plates and the op50 plates were removed and then placed back into the incubator. The op500 was the negative control plate wtih 80 and 300 progeny and unc-22 had around 50. (reference plates) Mating in the RNAi plates Dpy (60) x wildtype (15) and unc( 70) x wildtype ( 8) progeny the phenotype was movement and it seemed similar phenotype within the progeny. Can you help me understand the results and what is happening genetically in the experiment ? Are the for dyp-15 x unc-22 mating cross and unc-22 x eri-1 wildtype C. elegans. Based on the data: Briefly discuss the results of the RNAi experiment. How closely did the RNAi-treated worms phenocopy the mutant worms? Was the gene knockdown 100% effective? Was the RNAi treatmentequally effective for the two genes? If not, discuss this result and why that could occured. Please be specific and detailed

nothing is missing now.you just have to tell what are expected observations.AS WE HAVE TO DO EXPERIMENT AFTER BUT FIRST WRITE THAT WHAT WILL BE EXPECTED FROM THE RESULTS?WE WILL GET PHOTOS WHEN WE PERFORM BUT THEN WE WILL COMPARE WHETHER WHAT WE EXPECTED IS WHAT WE GOT OR IS THE ACTUAL RESULT DIFFERENT.
INTRODUCTION
For acquiring food material, bacteria excrete extracellular enzymes that split larger molecules into smaller units.For example, amylase and cellulase degrade polysaccharides such as starch and cellulose into simple sugars, which are then transported into the cell for metabolic purposes. A variety of proteases degrade protein molecules such as gelatin and casein into amino acids, and lipases split fats into fatty
acids and glycerol. These physiological or biochemical characteristics are used for isolation and identification of bacteria in conjunction with other morphological and cultural methods. In this exercise, you will perform tests related to hydrolytic reactions.
Starch hydrolysisâSince many bacteria are capable of hydrolyzing starch, this test has a fairly wide application. Bacteria that hydrolyze starch produce amylases that yield molecules of maltose, glucose, and dextrins.
Casein hydrolysisâCasein is the predominant protein in milk and its presence causes milk to have its characteristic white color. Many bacteria produce proteases that degrade proteins into peptides and individual amino acids in a process called proteolysis.

MATERIALS
ï·
Nutrient broth cultures of Bacillus subtilis, Escherichia coli andStaphylococcus epidermidis
ï·
Starch agar culture plate
ï·
Casein agar culture plate
ï·
Gramâs iodin

1.Label and streak in a straight line with all the 3 cultures on the starch agar plate ANDon the casein agar plate.

2.Incubate the plates at 37oC for 24-48 hours.

Week 2 âStarch Hydrolysis

1.Add several drops of Gramâs iodine over the growth on the medium. Rotate and tilt the plate gently to spread the iodine. Iodine is a starch indicator and when it comes in contact with a medium of starch, it turns blue.

2.If starch is hydrolyzed and no longer present, the medium will have a clear zone next to the growth.

3. Take a photograph of your plate against a white background. (If you do not have a phone equipped with a camera, ask a lab mate to take a picture for you and have the picture electronically sent to you.)

Week 2 âCasein Hydrolysis

1.Examine the streaks on the casein agar plates. A clear zone adjacent to the growth is evidence of casein (protein) hydrolysis. Discussion Amongst other points, you should address in the discussion whether the observations you made were expected. (You will need to do research to answer address this issue. Make sure you include proper citations?

What is expected observation for it with reference if possible