Dan:Notebook/HiResChrPaint/2013-4-10
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LambdaExo Digestion Gel Quantification
Urea gel to quantify Lambda Exonuclease digestion from 4/5/2013
Urea Gel Protocol
Prerun gel for at least 20 minutes at 250V If the gel stops running before samples are ready just repeat Set up samples according to table below
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 0.5L | 1L | 2L | V4S1 | V4S2 | V4S3 | V4S4 | V6S1 | V6S2 | V6S3 | V6S4 | 0.5L |
uL Sample | 0.5 | 1 | 2 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 | 0.5 |
uL ddH20 | 5.5 | 5 | 4 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 | 5.5 |
uL 2x Urea Buffer | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 | 6 |
Ran gel at 250V for 25 minutes
Gel Images
File:LambdaExo Digest 041013.jpg File:LambdaExo Digest Sampleid 041013.jpg
Quantification
Name | Sample | Volume | Minus Back | ng | sample ng | ng/uL |
INT*mm2 | ||||||
U1 | 100 bp 0.5 uL | 3734.900111 | 1179.300035 | 1.25 | ||
U2 | 200 bp 0.5 uL | 5710.70017 | 3216.350096 | 2.5 | ||
U3 | 100 bp 1 uL | 5355.74016 | 2704.900081 | 2.5 | ||
U4 | 200 bp 1 uL | 9250.050276 | 6494.800194 | 5 | ||
U5 | 100 bp 2 uL | 7312.240218 | 4959.430148 | 5 | ||
U6 | 200 bp 2 uL | 14064.53042 | 9710.270289 | 10 | ||
U7 | 100 bp 0.5 uL | 2633.210078 | 832.4200248 | 1.25 | ||
U8 | 200 bp 0.5 uL | 2938.250088 | 1623.040048 | 2.5 | ||
U9 | V4S1 | 8283.920247 | 2663.760079 | 2.60 | 5.19 | |
U10 | V4S2 | 29179.31087 | 15496.58046 | 14.15 | 28.29 | |
U11 | V4S3 | 26099.06078 | 14960.41045 | 13.66 | 27.33 | |
U12 | V4S4 | 10789.25032 | 5057.100151 | 4.75 | 9.50 | |
U13 | V6S2 | 18800.71056 | 12526.75037 | 11.47 | 22.95 | |
U14 | V6S3 | 21088.09063 | 13177.76039 | 12.06 | 24.12 | |
U15 | V6S4 | 22420.46067 | 15236.43045 | 13.91 | 27.82 | |
U16 | 2555.600076 | |||||
U17 | 2494.350074 | |||||
U18 | 2650.840079 | |||||
U19 | 2755.250082 | |||||
U20 | 2352.81007 | |||||
U21 | 4354.26013 | |||||
U22 | 1800.790054 | |||||
U23 | 1315.210039 | |||||
U24 | 5620.160167 | |||||
U25 | 13682.73041 | |||||
U26 | 11138.65033 | |||||
U27 | 5732.150171 | |||||
U28 | 6273.960187 | |||||
U29 | 7910.330236 | |||||
U30 | 7184.030214 |