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=Probe Prep Test 2 ([[Daniel:Notebook/HiResChrPaint/2013-5-10|Started 5/10/2013]])= [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] ==Ethanol Precipitation== 7. Centrifuged 10000 rpm for 30 minutes at 4C 8. Removed supernatant added 750 uL 75% EtOH 9. Centrifuged 10000 rpm for 12 minutes at 4C 10. Removed supernatant, dried in hood 11. Resuspended with 50 uL 12. [[Daniel:Protocols/Qiaquick_Column|Column Purification]], eluted with 50 uL 13. measured concentration in nanodrop ===Nanodrop Results=== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt;font-weight:bold" align="center" valign="bottom" | width="65" height="15" | Sample | width="65" | V4A | width="65" | V4B | width="65" | V4C | width="65" | V4D | width="65" | V6A | width="65" | V6B | width="65" | V6C | width="65" | V6D |- style="font-size:12pt" align="center" valign="bottom" |style="font-weight:bold" height="15" | ng/uL | align="center" | 127.5 | align="center" | 156.3 | align="center" | 130.2 | align="center" | 130.6 | align="center" | 142.7 | align="center" | 138.9 | align="center" | 146.4 | align="center" | 133.4 |- style="font-size:12pt" align="center" valign="bottom" |style="font-weight:bold" height="15" | ng | align="center" | 5100 | align="center" | 6252 | align="center" | 5208 | align="center" | 5224 | align="center" | 5708 | align="center" | 5556 | align="center" | 5856 | align="center" | 5336 |- style="font-size:12pt" align="center" valign="bottom" |style="font-weight:bold" height="15" | ug in 4 sets | align="center" | | align="center" | | align="center" | | align="center" | 21.8 | align="center" | | align="center" | | align="center" | | align="center" | 22.5 |} ==DpnII Digestion== [[Daniel:Reagents/DpnII|Enzyme Info]] 1. Prepared samples according to table *DpnII-Lot 0091201; Buffer 0031105 {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt;font-weight:bold" align="center" | width="65" height="40" | Sample | width="65" | V4A | width="65" | V4B | width="65" | V4C | width="65" | V4D | width="65" | V6A | width="65" | V6B | width="65" | V6C | width="65" | V6D |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | ng/uL | align="center" | 127.5 | align="center" | 156.3 | align="center" | 130.2 | align="center" | 130.6 | align="center" | 142.7 | align="center" | 138.9 | align="center" | 146.4 | align="center" | 133.4 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | ng | align="center" | 5100 | align="center" | 6252 | align="center" | 5208 | align="center" | 5224 | align="center" | 5708 | align="center" | 5556 | align="center" | 5856 | align="center" | 5336 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | uL | align="center" | 35 | align="center" | 35 | align="center" | 35 | align="center" | 35 | align="center" | 35 | align="center" | 35 | align="center" | 35 | align="center" | 35 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="35" | uL 10x buffer | align="center" | 10 | align="center" | 10 | align="center" | 10 | align="center" | 10 | align="center" | 10 | align="center" | 10 | align="center" | 10 | align="center" | 10 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="32" | uL DpnII (50U/uL) | align="center" | 2 | align="center" | 2 | align="center" | 2 | align="center" | 2 | align="center" | 2 | align="center" | 2 | align="center" | 2 | align="center" | 2 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | uL nf H20 | align="center" | 53 | align="center" | 53 | align="center" | 53 | align="center" | 53 | align="center" | 53 | align="center" | 53 | align="center" | 53 | align="center" | 53 |- style="font-size:12pt" align="center" |style="font-weight:bold" height="15" | Total | align="center" | 100 | align="center" | 100 | align="center" | 100 | align="center" | 100 | align="center" | 100 | align="center" | 100 | align="center" | 100 | align="center" | 100 |} 2. Incubated 16 hours at 37C, followed by 20 minutes at 65C; Continued on [[Daniel:Notebook/HiResChrPaint/2013-5-12|5/12/2013]]
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