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\author{woozing}
\pdfinfo{
  /Title (plant-and-tissue-culture-c3-media-preparation.pdf)
  /Creator (Cheatography)
  /Author (woozing)
  /Subject (PLANT \& TISSUE CULTURE - C3 (Media Preparation) Cheat Sheet)
}

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    {\parbox{\dimexpr\textwidth-2\fboxsep\relax}{\noindent
        \hspace*{-6pt}\includegraphics[width=5.8cm]{/web/www.cheatography.com/public/images/cheatography_logo.pdf}}
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    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{PLANT \& TISSUE CULTURE - C3 (Media Preparation) Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{woozing} via \textcolor{DarkBackground}{\uline{cheatography.com/146689/cs/31795/}}}
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  \mymulticolumn{2}{p{5.377cm}}{\bf\textcolor{white}{Cheatographer}}  \\
  \vspace{-2pt}woozing \\
  \uline{cheatography.com/woozing} \\
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Not Yet Published.\\
   Updated 22nd April, 2022.\\
   Page {\thepage} of \pageref{LastPage}.
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  %\includegraphics[width=48px,height=48px]{dave.jpeg}
  Measure your website readability!\\
  www.readability-score.com
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Culture Medium Components}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650599804_Screenshot 2022-04-22 115315.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{3.92 cm} x{4.08 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Media for Plant Cultures}}  \tn
% Row 0
\SetRowColor{LightBackground}
1. Macronutrients (mM concentrations) & N, P, K, Ca, Mg, S \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
2. Micronutrients (µm concentrations) & Fe, B, Cu, Mn, Zn, Mo, I, Co \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
3. Carbon Source & sucrose, glucose, other sugar \tn 
% Row Count 6 (+ 2)
% Row 3
\SetRowColor{white}
4. Vitamins & thiamine, biotin, pantothenic acid, nicotinic acid, pyridoxine, folic acid, ascorbic acid, tocopherol, myo-inositol \tn 
% Row Count 12 (+ 6)
% Row 4
\SetRowColor{LightBackground}
5. Complex Organic Supplements & coconut water, banana powder, yeast extract, peptone, potato homogenate \tn 
% Row Count 16 (+ 4)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Macronutrients}}  \tn
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\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650600524_Screenshot 2022-04-22 120823.png}}} \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Carbon and Energy Source}}  \tn
% Row 0
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\mymulticolumn{1}{x{8.4cm}}{Every living organism needs to have a source of energy in order to complete all the vital processes within the organism, and therefore each medium needs sugars as a source of carbon and energy.} \tn 
% Row Count 4 (+ 4)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{The preferred carbohydrate in plant cell culture media is sucrose.} \tn 
% Row Count 6 (+ 2)
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Vitamins}}  \tn
% Row 0
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\mymulticolumn{1}{x{8.4cm}}{Vitamins work as an assistant in enzymatic systems.} \tn 
% Row Count 2 (+ 2)
% Row 1
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\mymulticolumn{1}{x{8.4cm}}{They are required in very small amounts.} \tn 
% Row Count 3 (+ 1)
% Row 2
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\mymulticolumn{1}{x{8.4cm}}{Thiamine (B1), is more commonly used in plant tissue cultures and other vitamins such as nicotinic acid, pyridoxine (B6) etc.} \tn 
% Row Count 6 (+ 3)
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% Row 0
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\mymulticolumn{1}{x{8.4cm}}{-involved in the regulation of growth and organized organ development of plant tissues directly or indirectly} \tn 
% Row Count 3 (+ 3)
% Row 1
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\mymulticolumn{1}{x{8.4cm}}{-interactions of auxin and cytokinin are considered to be the most important regulation to induce organ development in the cultured tissues} \tn 
% Row Count 6 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-requirement of hormones (gibberellin, abscisic acid, ethylene) which will help to induce the developmental response in cultures depends on the type of explant and species to be cultured} \tn 
% Row Count 10 (+ 4)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-addition of growth regulators in the culture media also depends on the goal of the culturing process} \tn 
% Row Count 13 (+ 3)
% Row 4
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\mymulticolumn{1}{x{8.4cm}}{-i.e. gibberellin, ethylene, and abscisic acid are not required for organ development or cell proliferation in culture} \tn 
% Row Count 16 (+ 3)
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Five Classes of Plant Hormones/Growth Regulators}}  \tn
% Row 0
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\mymulticolumn{1}{x{8.4cm}}{1. Auxin} \tn 
% Row Count 1 (+ 1)
% Row 1
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\mymulticolumn{1}{x{8.4cm}}{2. Cytokinin} \tn 
% Row Count 2 (+ 1)
% Row 2
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\mymulticolumn{1}{x{8.4cm}}{3. Gibberellin} \tn 
% Row Count 3 (+ 1)
% Row 3
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% Row Count 4 (+ 1)
% Row 4
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% Row Count 5 (+ 1)
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{1. Auxins}}  \tn
% Row 0
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\mymulticolumn{2}{x{8.4cm}}{-Naturally occurring: indole-3-acetic acid (IAA)} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Synthetic: 2,4-dichlorophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (NAA), idole-3-butyric acid (IBA)} \tn 
% Row Count 4 (+ 3)
% Row 2
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\seqsplit{-Functions:} & a) Cell division and differentiation (with cytokinin) \tn 
% Row Count 6 (+ 2)
% Row 3
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 & b) Shoot and root apical dominance \tn 
% Row Count 8 (+ 2)
% Row 4
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 & c) Parthenocarpy in some species \tn 
% Row Count 10 (+ 2)
% Row 5
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% Row Count 12 (+ 2)
% Row 6
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 & e) At high concentration will kill plant as herbicide \tn 
% Row Count 14 (+ 2)
% Row 7
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{*Parthenocarpy is the natural or artificially induced production of fruit without fertilization of ovules, which makes the fruit seedless} \tn 
% Row Count 17 (+ 3)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{*Abscission is the shedding of various parts of an organism, such as a plant dropping a leaf, fruit, flower} \tn 
% Row Count 20 (+ 3)
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{2. Cytokinin}}  \tn
% Row 0
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\mymulticolumn{2}{x{8.4cm}}{-Naturally occurring: zeatin} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Synthetic: kinetin, benzylaminopurine (BAP) and adenine} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Synthesis from adenine in root tips, embryos,  young fruits, leaves in all plants} \tn 
% Row Count 5 (+ 2)
% Row 3
\SetRowColor{white}
\seqsplit{-Functions:} & a) Growth and development (in combination with auxin) \tn 
% Row Count 7 (+ 2)
% Row 4
\SetRowColor{LightBackground}
 & b) Delay senescence (Plant senescence is the process of aging in plants) \tn 
% Row Count 10 (+ 3)
% Row 5
\SetRowColor{white}
 & c) Break apical dominance \tn 
% Row Count 11 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Collaboration of Auxin and Cytokinin}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650606110_Screenshot 2022-04-22 134103.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\begin{tabularx}{8.4cm}{x{1.6 cm} x{6.4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{3. Gibberellins}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Naturally occurring: gibberellic acid-3 (GA3), GA4, GA7 (more than 90 different GA recognized)} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Synthesized from mevalonate shoot and root apices, embryos, cotyledons, fruits, tubers} \tn 
% Row Count 4 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\seqsplit{-Function:} & a) stem elongation and flowering \tn 
% Row Count 6 (+ 2)
% Row 3
\SetRowColor{white}
 & b) effects on seed germination (breaking seed dormancy) \tn 
% Row Count 8 (+ 2)
% Row 4
\SetRowColor{LightBackground}
 & c) promotes cell division in combination with IAA \tn 
% Row Count 10 (+ 2)
% Row 5
\SetRowColor{white}
 & d) Improves fruit set, fruit growth, fruit maturation and fruit ripening \tn 
% Row Count 13 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 13 (+ 0)
% Row 7
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{In tissue culture, GA's (gibberellic acids) are supplemented in some operations and avoided in others.} \tn 
% Row Count 16 (+ 3)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-For example, the presence of GA's in media can inhibit organ development (root and shoot formation) and somatic embryogenesis.} \tn 
% Row Count 19 (+ 3)
% Row 9
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-However, gibberellic acids are necessary to induce normal callus growth. Similarly, gibberellic acids can inhibit the meristemoid initiation; the catch is the meristemoid initiation is required for the growth and development of organs that are already formed.} \tn 
% Row Count 25 (+ 6)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\begin{tabularx}{8.4cm}{x{1.6 cm} x{6.4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{4. Ethylene}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-naturally occurring gaseous hormone that plays a role in fruit ripening, senescence, and leaf abscission} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\seqsplit{-Function:} & a) wound responses \tn 
% Row Count 5 (+ 2)
% Row 2
\SetRowColor{LightBackground}
 & b) causes thickening of stems and leaf abscission (aging) \tn 
% Row Count 7 (+ 2)
% Row 3
\SetRowColor{white}
 & c) reduces adventitious shoot formation \tn 
% Row Count 9 (+ 2)
% Row 4
\SetRowColor{LightBackground}
 & d) control fruit ripening in climacteric fruit \tn 
% Row Count 11 (+ 2)
% Row 5
\SetRowColor{white}
 & e) inhibits the growth and development of the plants in the culture at a higher concentration; but enhances the responses of plants towards auxin at lower concentrations \tn 
% Row Count 17 (+ 6)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{*Silver nitrate (AgNO3) has anti-ethylene activity} \tn 
% Row Count 18 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{5. Abscisic acid}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-maintains bud and seed dormancy, inhibits cell wall acidification and slows cell elongation} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-promotes somatic embryogenesis at a lower concentration; but halts the developmental response of cultures at a higher concentration} \tn 
% Row Count 5 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Used in agriculture where seed dormancy is important} \tn 
% Row Count 7 (+ 2)
% Row 3
\SetRowColor{white}
\seqsplit{-Function:} & a) water stress response \tn 
% Row Count 9 (+ 2)
% Row 4
\SetRowColor{LightBackground}
 & b) seed protein synthesis \tn 
% Row Count 10 (+ 1)
% Row 5
\SetRowColor{white}
 & c) seed dormancy \tn 
% Row Count 11 (+ 1)
% Row 6
\SetRowColor{LightBackground}
 & d) seed germination (in combination with gibberellins) \tn 
% Row Count 13 (+ 2)
% Row 7
\SetRowColor{white}
 & e) enhance somatic embryogenesis \tn 
% Row Count 14 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\begin{tabularx}{8.4cm}{x{1.44 cm} x{6.56 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Support Matrices}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{1. Agar} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-commonly used gelling agent in plant tissue culture} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-mixture of polysaccharides derived from red algae} \tn 
% Row Count 4 (+ 1)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Agarose is often used when the impurities in agar are not desired, such as in protoplast and anther culture} \tn 
% Row Count 7 (+ 3)
% Row 4
\SetRowColor{LightBackground}
\seqsplit{-contain:} & a) 70\% agarose (gelling component) (polymer of alternating D-galactose and 3,6anhydrogalactose) \tn 
% Row Count 10 (+ 3)
% Row 5
\SetRowColor{white}
 & b) 30\% agaropectin (non-gelling fraction, polymer of sulphated D-galactose units) \tn 
% Row Count 13 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 13 (+ 0)
% Row 7
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{2. Gellan Gums} \tn 
% Row Count 14 (+ 1)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-gelling agent used for plant tissue and cell culture} \tn 
% Row Count 16 (+ 2)
% Row 9
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-produces a high transparent gel, which allows better observation (inspection of contamination) of root growth compared with conventional agar gel} \tn 
% Row Count 19 (+ 3)
% Row 10
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-polymers from glucose, glucoronic acid and rhamnose units} \tn 
% Row Count 21 (+ 2)
% Row 11
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-not only easier for root inspection, but allow better root growth than agar} \tn 
% Row Count 23 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{2 cm} x{6 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Antibiotics}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-plants are sensitive to many antibiotics} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-use of antibiotics to prevent contamination is uncommon in plant tissue culture} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 3 (+ 0)
% Row 3
\SetRowColor{white}
1. \seqsplit{Cefotaxime} & kill Agrobacterium but not the plant cell \tn 
% Row Count 5 (+ 2)
% Row 4
\SetRowColor{LightBackground}
2. Kanamycin & kill plant cell \tn 
% Row Count 7 (+ 2)
% Row 5
\SetRowColor{white}
 & -Transgenic plant carrying the kanamycin selectable marker would survive on medium containing kanamycin \tn 
% Row Count 11 (+ 4)
% Row 6
\SetRowColor{LightBackground}
 & -Non transgenic plant would not survive Plant Tissue Culture (PTC) \tn 
% Row Count 14 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Step-By-Step Guide to Prepare Medium}}  \tn
% Row 0
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\mymulticolumn{2}{x{8.4cm}}{Pre-Preparation} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
1. Prepare 1 L of MS agar medium with 1 mg/L 2,4-D & a) Prepare all the stock solutions \tn 
% Row Count 4 (+ 3)
% Row 2
\SetRowColor{LightBackground}
 & b) Measure 800 mL of deionised water into a 2 L beaker \tn 
% Row Count 7 (+ 3)
% Row 3
\SetRowColor{white}
 & c) Put in a magnetic stirrer and start stirring \tn 
% Row Count 10 (+ 3)
% Row 4
\SetRowColor{LightBackground}
 & d) Add 50 mL of MS macro, 5 mL of MS micro, MS ferum and MS vitamin \tn 
% Row Count 14 (+ 4)
% Row 5
\SetRowColor{white}
 & e) Weigh 30 g of sucrose \tn 
% Row Count 16 (+ 2)
% Row 6
\SetRowColor{LightBackground}
 & f) Add into the mixture, stir until dissolve \tn 
% Row Count 19 (+ 3)
% Row 7
\SetRowColor{white}
 & g) Add 200 μL of 2,4-D \tn 
% Row Count 21 (+ 2)
% Row 8
\SetRowColor{LightBackground}
 & h) Adjust pH to 5.5 \tn 
% Row Count 22 (+ 1)
% Row 9
\SetRowColor{white}
 & i) Top up medium to 1 L \tn 
% Row Count 24 (+ 2)
% Row 10
\SetRowColor{LightBackground}
2. Weigh 5 g of agar & 1a) Autoclave, then pour medium \tn 
% Row Count 26 (+ 2)
% Row 11
\SetRowColor{white}
 & 1b) Pour medium into a 2 L bottle. \tn 
% Row Count 28 (+ 2)
% Row 12
\SetRowColor{LightBackground}
 & 1c) Add agar into medium in the bottle. Swirl to mix \tn 
% Row Count 31 (+ 3)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Step-By-Step Guide to Prepare Medium (cont)}}  \tn
% Row 13
\SetRowColor{LightBackground}
 & 1d) Autoclave medium. Swirl after autoclaving \tn 
% Row Count 3 (+ 3)
% Row 14
\SetRowColor{white}
 & 1e) Allow medium to cool \tn 
% Row Count 5 (+ 2)
% Row 15
\SetRowColor{LightBackground}
 & 1f) Pour medium into containers in a laminar air flow cabinet \tn 
% Row Count 9 (+ 4)
% Row 16
\SetRowColor{white}
 & 2) Dispense, then autoclave \tn 
% Row Count 11 (+ 2)
% Row 17
\SetRowColor{LightBackground}
 & 2a) Add agar into medium in the 2 L beaker \tn 
% Row Count 14 (+ 3)
% Row 18
\SetRowColor{white}
 & 2b) Dissolve agar in a microwave oven \tn 
% Row Count 16 (+ 2)
% Row 19
\SetRowColor{LightBackground}
 & 2c) Dispense medium into containers \tn 
% Row Count 18 (+ 2)
% Row 20
\SetRowColor{white}
 & 2d) Autoclave all the containers with medium \tn 
% Row Count 21 (+ 3)
% Row 21
\SetRowColor{LightBackground}
 & 2e) Allow to cool \tn 
% Row Count 22 (+ 1)
% Row 22
\SetRowColor{white}
3. Allow medium to cool and incubate for at least 3 days to 5 days before use. & -if contaminants present, it would appear during this period \tn 
% Row Count 26 (+ 4)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}


% That's all folks
\end{multicols*}

\end{document}