Noi/NOTES/2014-4-14

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Revision as of 16:08, 15 April 2014 by >Noi (Created page with "= gDNA extraction of cell cultures (Song' samples) = : '''Sample info.''' :: - Three vials for day 0 ESCs (ESCd0.1, ESCd0.2,ESCd0.3) :: - Three vials for definitive endoderm c...")
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gDNA extraction of cell cultures (Song' samples)

Sample info.
- Three vials for day 0 ESCs (ESCd0.1, ESCd0.2,ESCd0.3)
- Three vials for definitive endoderm cells (DEd3.1,DEd3.2,DEd3.3)
- Two vials for naive cells. (naiveESC.1, naiveESC.2)
- Extract gDNA using DNeasy Blood & Tissue Kit (QIAGEN) with RNaseA treatment and followed the instruction of the kit
- Elute DNA with 200ul AE buffer (single elution)
- Measure concentration with Nanodrop (ND) and Qubit dsDNA HS assay

Qubit dsDNA HS assay and Nanodrop (ND) quantification

Sample Conc. in the Qubit Unit uL used Dilution Sample conc. (ng/ul) Yields in 200ul (ug) conc. By ND (ng/ul) 260/280 260/230
ESCd0.1 201 ng/mL 1 200 40.30 8.06 53.15 1.92 1.88
ESCd0.2 280 ng/mL 1 200 56.00 11.20 76.53 1.88 1.81
ESCd0.3 430 ng/mL 1 200 86.00 17.20 107.13 1.91 1.94
DEd3.1 170 ng/mL 1 200 34.00 6.80 44.91 1.91 2.07
DEd3.2 188 ng/mL 1 200 37.70 7.54 47.82 1.91 2.02
DEd3.3 129 ng/mL 1 200 25.70 5.14 34.79 1.84 1.85
naiveESC.1 132 ng/mL 1 200 26.30 5.26 39.75 1.86 1.55
naiveESC.2 150 ng/mL 1 200 30.00 6.00 44.82 1.86 1.52
- For accuracy, the concentration will be based on Qubit measurement