User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/26

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AMPure Cleaned PCR Product

07April10 Ethanol Cleaned Product (~30ul) - Consumed 10ul of remaining Product

- Used Standard Agencourt AMPureXP Protocol http://www.beckmangenomics.com/products/dna_purification_and_cleanup/agencourt_ampure_xp.html

    • Final Elution step, only could retrieve ~30ul cleanly, minimal bead pick up.
Sample ID Nucleic Acid Conc. Unit A260 A280 260/280 260/230 Sample Type
lco/hco 10.5ng/µl0.010.0051.913.78DNA
lco/hco 91ng/µl0.020.0161.262.17DNA
lco/hco 17-0.1ng/µl-0.003-0.0170.170.15DNA
lco/hco 251.5ng/µl0.030.0083.980.84DNA
lco/hco 331.2ng/µl0.0250.021.251.73DNA
lco/hco 410.7ng/µl0.014-0.014-11.66DNA
lco/hco 491.7ng/µl0.0350.0221.561.21DNA
lco/hco 571.2ng/µl0.0240.0131.821.07DNA
lco/hco 651.5ng/µl0.030.0281.081.25DNA
lco/hco 731.7ng/µl0.0340.0142.51.19DNA
' 10ul Ethanol Cleaned -> Total PCR Product Recovery
AMPure Cleaned = 30ul
lco/hco 19715
lco/hco 934030
lco/hco 1722310
lco/hco 2529945
lco/hco 3328736
lco/hco 4156421
lco/hco 4959751
lco/hco 5737036
lco/hco 6528345
lco/hco 7343151


PEG Cleaned PCR Product

Clean remaining 07April10 Ethanol Cleaned PCR Product ~18ul with PEG.

To make PEG: 3g PEG 8000 2.19g NaCl Bring up to 15ml with good water. Lasts 30 days

Protocol: 1. Add 1:1 volume of PEG to PCR product. 2. Vortex briefly. 3. Warm at 37degrees for 15minutes. 4. Microcentrifuge for 15minutes at 4000rpm (cold is best). 5. Pipette off supernatant. 6. Wash pellet with 100ul 80% cold ethanol. 7. Spin for 10minutes at 4000rpm. Cold is best. 8. Pipette off ethanol. 9. Repeat steps 6-8 once or twice. 10. Dry down. 11. Resuspend in 10ul good water. 12. NanoDrop

1 0.2 ng/µl 0.004 -0.011 -0.38 0.17 DNA 50
90.4ng/µl0.007-0.013-0.530.71DNA50
170.5ng/µl0.01-0.016-0.60.3DNA50
252.8ng/µl0.0570.0232.50.37DNA50
330.4ng/µl0.008-0.011-0.710.89DNA50
411.5ng/µl0.030-1118.820.44DNA50
494.1ng/µl0.0820.0352.320.96DNA50
572.7ng/µl0.0540.02720.98DNA50
650.9ng/µl0.0180.00125.440.54DNA50
731.9ng/µl0.0390.0057.240.87DNA50
761.8ng/µl0.035-0.003-10.140.61DNA50
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