The species of plant that has been used in this experiment is papaya leaf or Carica Papaya leaf. The leaf is surface sterilized by placing in a 25 ml of 70% ethanol in a sterile tube and then followed by 10% Clorox that incubate for 2-5 minutes. This is because acidified alcohol is more effective as a disinfectant (Dodds & Roberts, 1990). After that, rinse with sterile distilled water three times. The tools such as forceps and scalpel would be used for all transfers of the leaf. Therefore, forceps tips should be dipped in ethanol and passed through flame which is Bunsen burner to prevent contamination of microorganism (Agrios, 2005).
Next, 15 disks from the papaya leaf were cut out using sterile forceps and scalpel and collected in an empty sterile Petri dish. After that, take 5 disks (one by one) and place them on first Petri dish containing co-cultivation media. The first Petri dish is labelled as a control for this experiment. Co-cultivation media are contains Murashige and Skoogs Basal Salts, Kinetin, BAP and agar.
Agrobacterium culture (grown overnight) is poured into the remaining disks on the Petri dish without media. Then gently swirl the plate and incubate for 5-10 minutes. This step is required to make sure that the leaf is fully covered by Agrobacterium culture. After that, the remaining leaf disks are transferred (5 each) from bacterial solution to the second and third Petri dish that containing co-cultivation media. Each of leaf is touched briefly on a sterile filter paper to remove excess liquid.
While handling this experiment, the culture dish must remain sterile. The culture dish should always covered and only open when to add a disk. This is because it can prevent the culture from contamination. The plates were...
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...he band produce was faint compared to other groups band. Faint band produced might be due to several causes. It might cause from the small amount of DNA used during electrophoresis. A low concentration might be due to volume added per well width. It can also be due to the degradation of DNA or the DNA was electrophoresed off the gel. The denaturation of DNA might also result in faint band. In order to avoid getting faint band in the future experiment, precaution step must be taken. The small amount of DNA can be solved by increasing the amount of DNA and avoid nuclease contamination of DNA markers. Use lower voltage, higher percentage gel or less time when electrophorese. To avoid denatured DNA, do not heat DA markers prior to electrophoresis. TE or buffer containing 20mM NaCl can be used to dilute the markers (Troubleshooting guide for the electrophoresis, n.d.).
Two members of the group were instructed to visit the laboratory each day of the experiment to water and measure the plants (Handout 1). The measurements that were preformed were to be precise and accurate by the group by organizing a standardized way to measure the plants. The plants were measured from the level of the soil, which was flat throughout all the cups, to the tip of the apical meristems. The leaves were not considered. The watering of the plants took place nearly everyday, except for the times the lab was closed. Respective of cup label, the appropriate drop of solution was added to the plant, at the very tip of the apical meristems.
The first day an unknown sample was assigned to each group of students. The first test applied was a gram stain to test for gram positive or gram-negative bacteria. The morphology of the two types of bacteria was viewed under the microscope and recorded. Then the sample was put on agar plates using the quadrant streak method for isolation. There were three agar plates; one was incubated at room temperature, the second at 30 degrees Celsius, and the third at 37 degrees Celsius. By placing each plate at a different temperature optimal growth temperature can be predicted for both species of bacteria.
The next day, 100 µL of an overnight culture of Salmonella growth will be transfer onto the center of a Petri plate containing tryptic soy agar where taken out from refrigerator. Sprea...
After the addition of the media, we insert an aeration tube inside and cover the lid with a cotton plug and start giving them aeration. This preparation has to be put on for 3 days under proper sunlight and 25-30 degree Celsius to observe if the culture is healthy/ potent or not depending on the color each culture portrays (The nanochloropsis culture should have a grass-green color to be seen as potent and the isochrysis culture should have a dark brown color to be seen as potent), if the colors seem dull and light, then that might mean that the culture is impotent.
After that bolt the slide dry with bibulous paper. After that examine the slide under the oil immersion lens. After determining the Gram stain reaction, 18 specific biochemical tests were performed for further analysis. The way of biochemical test was different but need to incubate at 37 degree Celsius. Phenol red lactose, phenol red dextrose, phenol red sucrose, methyl red, voges-proskauer, citrate utilization test, urea hydrolysis, and nitrate reduction are the media which are in test tube as liquid. Which were use Inoculating loop to deep tube with assigned bacteria. Triple sugar iron, and citrate utilization are in slant and we use Inoculating needle to deep tube with assigned bacteria. Again, starch hydrolysis, casein hydrolysis, lipid hydrolysis, oxidase test and catalase test were test from agar plate which use inoculating loop. For those media, which were in agar plate, inoculate with loop by making a line streak down the center of the agar. After that all 18 medias need to incubate at 37 degree Celsius for 48 hours. After 48 hours, some media show bacteria’s characteristics and morphology without adding extra reagent. Some media need reagent to examine morphology and physiology. Experiment such as MRVP needs methyl red, Barritt’s A, and Barritt’s
The materials that were used in this lab included a 100/1000 μL micropipetter, 4 agar plates with pre-poured LB broth , ice bath, hot water bath, micro centrifuge tubes, sterile loops, UV light, as well as pGLO plasmids and E.coli.
Methods: 4.5 mL E.Coli EDTA suspension was pipetted into a conical tube. After this 0,25 mL lysosome solution was put inside the same tube. Both were incubated at 37°C for so minutes. Once out of the incubator, 0.5 mL of 10% SDS was added. In order to ensure a good mixing of the liquids the tube was inverted continuously for several minutes. Incubation occurred once more but this time at 60°C for 10 minutes. Once cold down to room
The first step in conducting the experiment was setting up the benchtop following sterile technique. A solution of 30% ethanol and 70% water was used to sterilize the field, and all dishes were put into an autoclave. The next step was putting the bacterial culture onto different nutrient plates using materials from Bio-Rad Laboratories’ pGLO Bacterial Transformation kit. First, 250흻l of the transformation solution was put into two microtest tubes, one labeled +pGLO (the tube administered the plasmid) and the other labeled -pGLO (the tube
Summary Paragraphs: In three dishes, paper was placed in the bottom to absorb the moisture. Twenty-five spinach seeds were added to each using tweezers. Two milliliters of different solutions were then added to each. The solutions were deionized water, 1 mL water and 1 mL vinegar, and 1 mL water and 1 mL Borax. The water was used to form a neutral environment, the vinegar to form an acidic environment, and the Borax to form a basic environment. The petri dishes were then labeled, taped shut, and placed on the windowsill for seven days.
Seeds are placed in transparent petri dishes at the same time and exposed to the same light level.
The purpose of this lab was to extract chlorophyll and carotenoid pigments from fresh spinach leaves and separate and analyze these pigments using column chromatography and thin layer chromatography. Acetone was used as a polar solvent to dissolve the more polar pigments first (Xanthophylls, chlorophylls), while hexane was used as a nonpolar solvent to dissolve the more nonpolar pigments such as the carotenes. In addition to being used as the polar solvent, acetone was used to remove the spinach components that were not pigments such as cellulose which is insoluble. The column chromatography worked by eluting the nonpolar carotene pigments first because the alumina is polar and doesn’t absorb the nonpolar carotene. The polar components such
For roughly 30 years the Puna District of Hawaii had been a safe haven from the devastating effects of the papaya ringspot virus (PRSV). Suddenly, in the early nineties, 95% of Hawaii's second largest crop became threatened when PRSV arrived in the Puna District (Gonsalves et al. 1998). Researchers at the University of Hawaii and Cornell University had anticipated that this day would come, so in preparation they began developing a PRSV resistant papaya using newly developed genetic engineering techniques. The initial fruit of their labors was "SunUp", a transgenic papaya created using biolistics to insert the coat protein genes from PRSV into a non-transgenic variety of papaya known as "Sunset". As a result of the insertion, SunUp has resistance to PRSV. The next step was "Rainbow", an F1-hybrid derived by crossing the transgenic SunUp papaya with the non-transgenic "Kapoho" papaya. Rainbow combines the PRSV resistance of its transgenic parent with the yellow flesh, preferred by consumers, of its non-transgenic parent.
...ince, there is a need to use for advanced novel methods of culturing plant to furnish new means for quickly propagating,conserving of endangered species and also introducing exotic plants. The production of high quality planting material of exotic nature propagated from vegetative parts through tissue culture has created new opportunities in global trading. The exotic plants are advantageous for farmers;growers; nursery owners & rural employment. As exotic plants are restricted to their natural environment; the main benefit of tissue culture technology lies on production of high quality & uniform planting material that can be multiplied on a year round basis. The plant selected for such purpose is Stevia rabuadiana Bertoni. Objectives of study:
Tissue culture allows for the clonal propagation of plant (production of multiple copies of the same genotype).
After harvesting of crop, leaves are totally removed from roots by sharp knife in field. These leaves are used a fodder for cattle or left in field for green manuring. According to some farmers, it increases milk production of