Daniel:Notebook/HiResChrPaint/2013-8-9

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S2 Probe Prep (ULS; Started 8/8/2013)[edit]

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DpnII Digestion (Started 8/8/2013)[edit]

3. Column purification, elute with 30 uL EB
4. Measure in Nanodrop

Nanodrop Results[edit]

Sample V4S2AB V4S2CD V6S2A V6S2B V6S2C V6S2D
ng/uL 109.3 122.9 115.4 117 121 111.1
ug in 30 uL 3.3 3.7 3.5 3.5 3.6 3.3
Total ug   7.0       13.9

Gel Image

File:2013-8-9-DpnIIDigest.jpg

Lambda Exonuclease Digestion[edit]

1. Reaction Table
Sample V4S1C V4S1D V6S1A V6S1B V6S1C V6S1D
ng/uL 109.3 122.9 115.4 117 121 111.1
ug in 25 uL 2.7 3.1 2.9 2.9 3.0 2.8
uL added 25 25 25 25 25 25
uL 10x buffer 5 5 5 5 5 5
uL Lambda Exonuclease (5U/uL) 3 3 3 3 3 3
uL nfH2O 17 17 17 17 17 17
total 50 50 50 50 50 50
2. Incubate for 1 hour at 37C followed by 10 minute heat inactivation at 75C
3. Pool samples
4. ssDNA column purification, elute with 20 uL
5. Measure Concentration in Nanodrop

Nanodrop Results[edit]

  ng/uL ug in 20 uL
V4S2 94.7 1.9
V6S2 195.2 3.9

FISH[edit]

Alan and I prepped some slides for FISH. However, we accidentally used the wrong media for attaching to slides. The cells did not attach well even after about 4 hours. So Alan and I decided to try again Monday.