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文献和实验Human Embryonic Stem (ES) Cell Protocols――Matrigel Aliquoting and Plating
on ice to keep them cold**. Example calculation with Matrigel arriving at 13.841 mg/ml: 13.841mg/1ml = 2mg/x ml x = 0.145ml/tube Using good sterile technique, aliquot Matrigel to tubes, switching tips whenever Matrigel seems to be clogging
Preparation of total cellular RNA--提取总RNA方法
1) Harvest 1x105 to 108 cells. Wash 1x w/PBS and freeze in liquid N2 and store @ -70℃2) Resuspend each pellet in 1.5ml lysis buffer (300μl 5x Lysis, 75μl 200mM VR, 1.125ml H2 O3) Incubate on ice 5minutes and spin 20min at 8k at 4℃.4) Add equal
Dissociated Retina Immunohistochemistry
Solutions 100X trypsin 10 mg/ml in 1X PBS Sigma #T2271 100X soybean trypsin inhibitor 10 mg/ml in 1X PBS Sigma #T6522 1000X DNaseI 2mg/ml in HBSS Sigma #D4513 Explant Culture
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