TITLE: 2XYT Medium (Version 2-0) Haseloff Lab
AUTHORS: Fernando Guzman Chavez

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:center"><span style = "font-weight:bold;">Work instruction for 2xYT and 2xYTG medium preparation</span></div></div><div class = "text-block"><div class = "justify" style = "text-align:center">Following this recipe, you will obtain 1L of 2XYT medium</div></div><div class = "text-block"><div class = "justify" style = "text-align:center"></div></div><div class = "text-block"><div class = "justify" style = "text-align:justify"><span style = "font-weight:bold;">2X YT</span><span>is a standard growth medium used to cultivate</span><span style = "font-style:italic;">E. coli</span><span>. In cell-free system, this medium is commonly used to growth the cells.</span></div><style>
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							</style></div><div class = "text-block">NOTE :</div><div class = "text-block">The protocol described here is an adaptation from these papers:</div><div class = "text-block"><ul style = "list-style-type:disc;"><li style = "counter-reset:ol0;list-style-type:disc;"><span style = "font-weight:bold;">Adam D. Silverman</span><span>, Nancy Kelley-Loughnane, Julius B. Lucks, and Michael C. Jewett (2019).</span><span style = "font-style:italic;">Deconstructing Cell-Free Extract Preparation for in Vitro Activation of Transcriptional Genetic Circuitry</span><span>. ACS Synthetic Biology, 403-414. DOI: 10.1021/acssynbio.8b00430.</span></li><li style = "counter-reset:ol0;list-style-type:disc;"></li><li style = "counter-reset:ol0;list-style-type:disc;"><span style = "font-weight:bold;">Andriy Didovyk</span><span>, Taishi Tonooka, Lev Tsimring, and Jeff Hasty. (2017).</span><span style = "font-style:italic;">Rapid and Scalable Preparation of Bacterial Lysates for Cell-Free Gene Expression</span><span>. ACS Synthetic Biology, 2198-2208. DOI: 10.1021/acssynbio.7b00253.</span></li></ul></div><div class = "text-block"><ul style = "list-style-type:disc;"></ul></div></div>

[STEPS]
?. [2xYT and 2xYTG medium preparation]
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							For this media, following compounds have to be ready-to-use:2XYT powder1M K2HPO4solution1M KH2PO4solution
?. [2xYT and 2xYTG medium preparation]
1. Prepare for autoclavingVolumes indicated are sufficient for 1 L of 2XYT medium 2x YTPG MediumFor 400 mL 2xYTP liquid medium add 7.2g D-glucose (Mw 180.16 g/mol), for a final concentrationof 0.1MMix until glucose is dissolved and filter sterilize (0.22um filter)  ABCD1Compound,   Sum FormulaMW [g/mol]Required amount [g or mL]Concentration [g/L]22X YT powder(16g/L tryptone,10g/L yeast extract, 5g/L NaCln/a31 g3131M Potassium Phosphate Dibasic  [K2HPO4]174.1840 mL7[0.04M]41M Potassium Phosphate Monobasic [KH2PO4]136.0922 mL3[0.022M]
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							2x YTPG Medium
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							For 400 mL 2xYTP liquid medium add 7.2g D-glucose (Mw 180.16 g/mol), for a final concentrationof 0.1MMix until glucose is dissolved and filter sterilize (0.22um filter)
ABCD1Compound,   Sum FormulaMW [g/mol]Required amount [g or mL]Concentration [g/L]22X YT powder(16g/L tryptone,10g/L yeast extract, 5g/L NaCln/a31 g3131M Potassium Phosphate Dibasic  [K2HPO4]174.1840 mL7[0.04M]41M Potassium Phosphate Monobasic [KH2PO4]136.0922 mL3[0.022M]
?. [3. Literature/ References]
?. [4. Change history]
Protocol for 2xYTPG medium was included