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=May 4, 2010 (gDNA from Salk)= Sergio Ruiz Macías, PhD Gene Expression Lab (GEL-B) Stem Cell Facility The Salk Institute For Biological Studies 10010 North Torrey Pines Road 92037 La Jolla, CA, USA sruiz@salk.edu Tel. 858-453-4100, ext 1324 {| {{table}} border=1 | align="center" style="background:#f0f0f0;"|'''Fibroblast couple''' | align="center" style="background:#f0f0f0;"|'''concentration (ug/ul)''' | align="center" style="background:#f0f0f0;"|'''volume (ul)''' | align="center" style="background:#f0f0f0;"|'''passage number''' |- | HFF XF (Human Foreskin fibroblast Xeno free)||0.2||50||9|| |- | FiPS 3F #1||0.178||56||24|| |- | |||||| |- | keratinocyte couple|||||| |- | Keratinocyte MMTA (human foreskin keratinocytes MMTA)||0.5||20||5|| |- | KiPS 4F #8||0.42||23.8||28|| |- | |||||| |- | Astrocytes|||||| |- | human astrocytes||0.36||90||12|| |- | ASTiPS4F-4||0.51||90||10|| |- | ASTiPS4F-5||0.71||90||10|| |- | |||||| |- | HUVEC|||||| |- | Human HUVEC||1.12||90||2 |- | HUViPS4F-1||1.03||90||10 |- | HUViPS4F-3||0.47||90||15 |- | |} *genomic DNA was extracted using the QIAmp DNA Mini and Blood Extraction Kit from Qiagen. DNA is resuspended in AE buffer (10mM Tris Cl; 0.5mM EDTA) *priority: **Keratinocyte MMTA and KiPS 4F #8 ***Human HUVEC, HUViPS4F-1, HUViPS4F-3 ****HFF XF, FiPS 3F #1 ***** human astrocytes, ASTiPS4F-4, ASTiPS4F-5 **NOTE: **HUVEC samples had very low concentration after shearing and end-repair reactions. Thus we suspect the DNA concentration that Sergio has given might not be accurate, and we measured the concentration again with Nanodrop and Qubit. {| {{table}} | align="center" style="background:#f0f0f0;"|'''Sample ID''' | align="center" style="background:#f0f0f0;"|'''Nanodrop result (ng/ul)''' | align="center" style="background:#f0f0f0;"|'''Qubit result (ng/ul)''' | align="center" style="background:#f0f0f0;"|'''Average (ng/ul)''' |- | HUVEC||46.4||27.7||37.05 |- | HUV ips4F1||79.5||54||66.75 |- | HUV ips4F3||34.4||25.9||30.15 |- | |} *Measured the concentration of Astrocyte sets again, because the library construction results seemed to be problematic (see labnotes 8-7-2010) {| {{table}} border=1 | align="center" style="background:#f0f0f0;"|'''Sample ID''' | align="center" style="background:#f0f0f0;"|'''concentration (ng/ul)''' |- | Ast||23 |- | Ast iPS 4F4||37.4 |- | Ast iPS 4F5||51 |}
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