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for this paper, i just need the method section which is 3 pages. i upload 2 pictures which are the requirements for this paper. I also upload the notebook which has everything that we have done in the lab. we have done a proposal for this experiment which was not good because we changed the species (the sources is NOT good except 1 source and YOU HAVE TO USE IT). Also, i upload a VERY helpful PowerPoint about all details that should be included.NOTE:1. the only good source that you have to use is this :… its about how to make the coal slurry. you should not write the steps just mention that you follow this paper. 2. you should talk about how we will do the measurement even though we haven’t done that yet 3. FOLLOW THE POWER POINT CAREFULLY.4. cite the software that we are using (ANOVA:jmp) if you need any information or help just let me knowTHANK YOU SO MUCH


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Bio 321: Ecology Capstone 2019
Triad 12:
Dany Holbert
John Hageter
Modhi Alajmi
Lauren Dye
Effects of Coal Mine Runoff on Salix
(Willow) Species 94006 and 05x-281-043
Mailing addresses:
Life Science Building 53 Campus Dr, Morgantown, West Virginia 26506
Lauren Dye: [email protected]
Modhi Alajmi: [email protected]
John Hageter: [email protected]
Dany Holbert: [email protected]
Table of contents:
Solution Prep: 2/22/19………….……..Page 3
Willow Prep: 2/26/19….……………….Page 5
Willow Prep cont: 3/5/19……..……….Page 7
Treatment solution: 3/19/19…………..Page 8
Solution Prep: Week of 2/18-2/22/19
Goal: finalized project operations by switching to willow species, prep a 100L of 10%
Johnson’s solution
Background: Can’t obtain one of the soy species so had to switch to two willow (salix)
species: 94006 and 05x-281-043. Johnson solution is a standard solution for hydroponic
growth containing necessary nutrients.
-Reagents: johnson solution
-medium: hydroponics. 10% Johnson’s Solution (1957)
Observations: This week we finalized our project operations in creating 100L of 10%
strength Johnson’s solution as well as claiming our Salix plants for treatment which
includes 24 clones of 94006 and 05x-281-043. Later this week of early next week we
will be creating our stock treatment solution and organizing out bins to begin hydroponic
growth. We worked with Phil to create a procedure for making Johnson’s solution in the
case that we need more down the line. Michael and Christine let us know that if we
need to make more, the 219 prep labs have all the solutions that we need and we just
have to let them know before we do decide to make more. We may hold off a few days
to transplant the willow into our bins because there was a note on the potting room door
that said they were transferred to the growth chambers due to possible transplant shock
and we wouldn’t want to shock them anymore before treatment.
Ideas for future experiments: After preparing the solution, we will prepare the plants
(willow species).
Willow Prep: week of 2/25-3/1
Goal: Prep the willow species for the duration of the experiment and set up the
hydroponic system
Background: To ensure consistency in growth, each species will be watered
hydroponically and solutions (johnsons and runoff) will be allocated accordingly.
Hydroponics is used to grow plants with water that contains nutritions and other
essential minerals. This technique helps the plants to grow faster because they’re
getting all the needed nutrition directly from the water.
-Reagents: johnson solution, treatment solution (slurry) was not created yet but
will be later this week of 3/3
-medium: hydroponics
-2/26: both species weren’t ready, due to lack of sprouts preventing distinction
between genotypes, so couldn’t be transported into our system.
-Phil will genotype these cuttings in the wells when they sprout shoots
-No data
Data analysis: N/A
Ideas for future experiments:
-have a more organized system to categorizing genus of plants
*we are a little behind schedule due to the inability to differentiate between willow
genotypes (until budding occurs) so our schedule provided in the proposal is off.
-This week will solely be limited by the amount to time it takes to genotype the species.
Once these are grown enough to be identified, they will be placed into out Johnson’s
Solution for one week where they will acclimate to the solution and resist transfer shock
before treatment solution (synth. AMD) is added to our treatment groups.
Willow Prep continued: week of 3/4-3/8
-To make sure the plants grew enough so we can start the experiment
-transfer plants to our bins
Background: distinguishing genotypes based on leaf characteristics
-94006 has serrations on side
-05x will have longer and brighter leaves
-reagents: johnsons
-medium: hydroponics
-noticed: not enough budding (as of 3/ 5) to determine genotype so cant proceed
-no buds, can’t distinguish genotypes, can’t allocate species into respective bins to
proceed with experiment
Data analysis: N/A
Ideas for future experiment: more organized class setup
Treatment solution: week of 3/18-3/22
-transfer plants (after distinguishing genotypes) to respective bins
-make the treatment solution (coal slurry) and add it to the Johnson’s solution.
-change johnson solution
-record data: leading shoot height/leaf count and longest root measurement
-For the preparation of the coal slurry, we will be synthesized in lab based on the precharacterized ranges of heavy metallic ion concentrations (Tiwary, R. K., 2001). In this
paper, the slurry has a multiple metals dissolved in tap water at the respective
concentrations: calcium carbonate −600 to 8000 mg/L, sulfate 14.7–10,700 mg/L, iron
<0.01 to 4100 mg/L, manganese 0.02–136 mg/L, aluminum <0.01 to 128 mg/L and zinc <0.003 to 18.8 mg/L. How: -reagents: previously used johnson solution and new synthetic slurry (pic below) -medium: hydroponics. -noticed: there is enough budding to determine genotype so can proceed. Observations: Before adding the coal slurry, we noticed that the plants are growing at a rapid rate. Changed from 6 plants to 8 per bin, so relabeled to keep data organized. Data analysis: Ik_migkfNYc3k9Q8ptrFGY2gKMOrQ/edit#gid=827058653 Average shot length: 15.4671875 Average leaf number: 15 Average longest root length: 15.225 Ideas for future experiment: Do the same measurement (shot length, leaf number and longest root length) next week. 8 How to Write a Methods Section { • • Methods Due in Class, Week of March 26th See pages 42-47 in lab manual for more information on writing manuscript, and rubric General        Explain your methods in a way that is clear, but concise. Someone reading this should be able to replicate the experiment 2-4 pages Don’t be too specific. Just give what is needed to replicate and fully understand OK to say “We…”. This helps use active voice Even though you have not completed your experiments, write your methods as if you have Experimental Design, Why, What, When, Where, How Experimental Design  Include a table that shows your experimental design  Include both factors  All levels of both factors  The number of replicates Why were activities performed?  Link methods to objectives/hypotheses  clearly show which methods test which hypothesis  Use outside sources to justify the levels used for each factor  Also use outside sources if you used a previously published protocol What was done?  Use enough detail so that the experiment could be repeated  Avoid trivial details  Ask: does this really matter?  E.g. greenhouse number?, brand of equipment?, how you measure height?, etc.  You do need to include CITATIONS for equipment you used, naming the model and company (i.e. SAS JMP software, spadmeter, etc.) When were activities performed?  Only discuss this if it is important.  It may matter depending on the life cycle of your organism, or the time it takes for pots to dry, etc.  Ask: would it matter if someone performed this activity at a different time/season/year?  You may talk about the timeline such as:  “Germination occurred 3 days after planting”  “10 days after germination, we noticed/did…” Where were key activities performed?  Only if it is is important/relevant  In a greenhouse. In a growth chamber. How were key activities performed?  How were specific treatment applied?   This is where you want to be most specific! How did you measure each dependent variable?  Again, be specific but concise!  BUT: Don’t list materials or steps  Write in paragraph form with complete sentences Statistical Analysis   You haven’t done this yet, but you should be able to write this section. What does a 2-way ANOVA test?   What effects did you include in your ANOVA model?     Main effects (each factor on its own) Interactive effect of both factors What other test might you use to compare levels?   Relate back to your three objectives, and mention each dependent variable TUKEY KRAMER HSD? What is your significance threshold? What programs/software did you use?  Cite SAS JMP Include Figures or Diagrams, if helpful  Especially if you have a complicated research design set up.  Can also include pictures in your methods section IF it helps the reader understand Other Announcements  Intro outline also due by end of class week of March 26th  Intro FINAL draft due in class week of April 2nd ... Purchase answer to see full attachment

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