2013_12_11 NADH comparison on polystyrine and crystal plate
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NADH ε340 = 6220M-1cm-1
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NADHBuffer = 50mM phosphates, pH 7
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Molar Mass (g/mol)741.6
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single component stock conc. (mM)40polystyrine plate info:Falcon brand product # 35 1172
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solubility
50 mg/mL in water, more if basic
"microtest Flat Bottom" polystyrene individually package. Not tissue culture treated plate, 96 well.
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uL required11,512.00Order of operations:
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surplus factor21) Make buffer, read with pH probe. Says 7.06.
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uL to shoot for230242) Weigh 8.22 mg NADH & dissolve in 277.1 uL buffer. NADH bottle is from 2011.
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mg to shoot for682.9843) Add required uL of NADH solution to each polystyrine well
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mg actually weighed out8.2204) Add required uL of buffer to each polystyrine well
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uL buffer to add277.105) Read polystyrine plate for 10 min
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check: M of solution0.046) move solution to quartz plate and measure 10 min
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7) set up for a mult-hour run in the crystal plate. Unfortunately the software crashed and the data was not available. In the morning, A340 was equal in all wells, indicating all of the NADH had degraded.
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rowcoluL NADHuL buffertotal uLtrue_mM_NADHpath_cmtheoretical_OD_from_known_mM
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A901501500.000.38250.00
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A1001501500.000.38250.00
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A1111491500.270.38250.63
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A1211491500.270.38250.63
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B921481500.530.38251.27
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B1021481500.530.38251.27
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B1131471500.800.38251.90
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B1231471500.800.38251.90
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C941461501.070.38252.54
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C1041461501.070.38252.54
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C1151451501.330.38253.17
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C1251451501.330.38253.17
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D961441501.600.38253.81
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D1061441501.600.38253.81
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D1171431501.870.38254.44
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D1271431501.870.38254.44
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E981421502.130.38255.08
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E1081421502.130.38255.08
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E11101401502.670.38256.34
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E12101401502.670.38256.34
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F9121381503.200.38257.61
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F10121381503.200.38257.61
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F11161341504.270.382510.15
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F12161341504.270.382510.15
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G9201301505.330.382512.69
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G10201301505.330.382512.69
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RESULTS:
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uL.NADHuL.buffertotal.uLtrue_mM_NADHpath_cmtheoretical_OD_from_known_mMplate.materialmean.OD340
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1015015000.38250polystyrene0.174192105
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211491500.270.38250.63polystyrene0.737472368
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321481500.530.38251.27polystyrene1.252155263
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431471500.80.38251.9polystyrene1.748663158
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541461501.070.38252.54polystyrene2.308542105
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651451501.330.38253.17polystyrene2.644335526
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761441501.60.38253.81polystyrene2.932455263
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871431501.870.38254.44polystyrene3.083059211
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981421502.130.38255.08polystyrene3.194856579
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10101401502.670.38256.34polystyrene3.321723684
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11121381503.20.38257.61polystyrene3.432786842
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12161341504.270.382510.15polystyrene3.632619737
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13201301505.330.382512.69polystyrene3.647551316
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14015015000.38250quartz0.030160526
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1511491500.270.38250.63quartz0.482206579
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1621481500.530.38251.27quartz0.910155263
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1731471500.80.38251.9quartz1.301217105
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1841461501.070.38252.54quartz1.790414474
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1951451501.330.38253.17quartz2.123255263
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2061441501.60.38253.81quartz2.446368421
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2171431501.870.38254.44quartz2.699015789
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2281421502.130.38255.08quartz2.860985526
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23101401502.670.38256.34quartz3.078928947
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24121381503.20.38257.61quartz3.277535526
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25161341504.270.382510.15quartz3.531682895
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26201301505.330.382512.69quartz3.643409211
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Sheet1
Phosphate Buffer Solution
raw data -- added 2015/5/24