dharmafect 1 transfection protocol

0000550007 00000 n 0000021353 00000 n XFWS+U4NNjEzwpYxXhqrLYz+sPS+Egt8bU+LFUV5z/5Q/Xv+2bef8mJMVeVZvHnnYq7FU28rf8pB 0000569266 00000 n uuid:69DFC3FDBEA9DC1182F0D63CB98ED3BC y3M7yesGPK7S8imCxvG0KArev9hFPStaYqqw+SrCGS4la8u5nuWikLSNESjwXcupROnGBfszTNsa fc/8ZHxPNIY9rRI1ac7/AGk6bfsrhHJDxfzfDN/jjV3Z+Km9mMNSGAapFWTen3ZFVKOyup7uxku0 0000287407 00000 n 0000631884 00000 n 0000010870 00000 n /BV9/wBX68+9v+qmKu/wVff9X68+9v8Aqpirv8FX3/V+vPvb/qpirv8ABV9/1frz72/6qYq7/BV9 ANcfqOKUq85/8ofr3/bNvP8AkxJiryrN4887FXYqm3lb/lILH/jJ/wAatlWp/ui36T+9DJtfuZY9 2010-07-27T15:08:04-06:00 bH6p6saMLqJeTiduJK7UGxxVbfeftN05JWurK8V4EV5IgIGcVhu7l1+G5K1RLN+W/hxrXFVO/wDP xmp.iid:2E25F82EE84EDF11986A9631617289C4 +TcVdirsVZV+V/8Aynujf8Zm/wCTcmKvom/u/Ls2pvYanHbSTxW37yS5EVEiuSY/RrKeX7302qoF 0000021696 00000 n 0000021893 00000 n 0000021304 00000 n 0000490759 00000 n 0000487477 00000 n /SwTlBxa79SWb1G/0av+7SOIIFAO++Kpho/lHTdENsbKSUCzjaFFpCnJG2AlMMMZcqAACxrtU1O+ 3/pMg/6qYqlPmzzZ5VufKutW9vrWnzTTafdRxxx3ULO7tDIqqqrISSSdhir/AP/Z dsVWNJ5KnvtQsruDT47myLJdC5SBWZJY4J5H+Pcxn1kDE7V64qmAfy9PLKQ1lJNeBbeYgxM0wVea 0000639267 00000 n 0000608967 00000 n 5MQo98VQpEZnacCsgAA/enjQcqfDyoN/bFlQ70UGDfZYH5YsVkvWP/X/AIHFUq85/wDKH69/2zbz 0000446985 00000 n 0000512673 00000 n AQBIAAAAAQAB/+4AE0Fkb2JlAGSAAAAAAQUAArEj/9sAhAAKBwcHBwcKBwcKDgkJCQ4RDAsLDBEU 0000483952 00000 n As a result, non-viral, transfection methods of DNA delivery are of increasing interest. Synthetic DNA Delivery Systems is a comprehensive and current resource on DNA transfection. 0000024068 00000 n 850 0 obj <> endobj xref 850 452 0000000016 00000 n vq3Ey2cTW6LL6LAxskUZ29AcW/coea8XqOtNsVVZ/I1nc2ctpcahfSNPO109xygWUu9mdMNOFsqg This volume provides a wide range of protocols used in studying the nuclear envelope, with special attention to the experimental adjustments that may be required to successfully investigate this complex organelle in cells from various ... This comprehensive volume, a complement to the new three-volume treatment of cilia and flagella by King and Pazour, presents easy-to-follow protocols and detailed background information for researchers working with cilia and flagella. ... 0000023432 00000 n /lILH/jJ/wAatlWp/ui36T+9DJPMGoRQavcRNrkVkV4f6O9gJylUQ/3npNWvXrmod0lv6Vh/6mWD The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity. 0000522182 00000 n /qpirv0NH/1L0X/cTT/qpirv0NH/ANS9F/3E0/6qYq79DR/9S9F/3E0/6qYq79DR/wDUvRf9xNP+ 0000596035 00000 n WuWbalcMrvPJ6fGeLfgXP2uJH7Rr8RwlatJo9SuNZ0y7sjbLEsFtHGJgu5rc227Md6kCp37YE1Td Efficient non-viral transfection of THP-1 cells. 2. 0000021794 00000 n Dilute DharmaFECT 1 by adding 1.8 µL DharmaFECT 1 in 373.2 µL serum-free and antibiotic-free cell culture medium. xGWka7mNpD6ZMqrvKpB5FaU3xVQufPWl2t49i9vcvNHMkEgQRMF5JcyGQn1vsqLOSoHxbbKajFXQ Adobe InDesign 6.0 siRNA targeting renilla luciferase at different final concentrations ranging from 0.5 to 20 nM was co-transfected with renilla luciferase gene . Table 2 presents 0000023774 00000 n 0000514308 00000 n 0000020913 00000 n 0000544996 00000 n 0000017977 00000 n KtN0PyxVCkw/XvT9Ahyqu05QcW+2oTn3YcemKogxxip4L9wxV3px/wAi7+wxVI9R1pbW6ls/q6Mi 552KuxVNvK3/ACkFj/xk/wCNWyrU/wB0W/Sf3oZD5i0lbnWLic6I96X4fv1vFhDUjQf3ZU0pSmah 0000016945 00000 n While DharmaFECT transfection reagent 1 is the most all-purpose formulation, recommended for 80% of cell lines tested, DharmaFECT transfection reagent 2, 3, and 4 offer chemically distinct alternatives for optimization prior to 0000447903 00000 n MS/MS confirmation or relative quantification was performed to identify lipid species altered in experimental conditions versus untreated control cells. 0000511046 00000 n 0000558383 00000 n 0000542182 00000 n owJZ/wCc/wDlD9e/7Zt5/wAmJMVeVZvHnnYq7FU28rf8pBY/8ZP+NWyrU/3Rb9J/ehk2v6Y1xq1x 0000624464 00000 n 6. 0000539264 00000 n 0000020616 00000 n �}��΋��t���w�E�z8�9/J�,�b���n8���ߒ�JGq�^��z8�>]�Y��7*=ꓜ��dgdN�?�٢ѡE�^�[+S�Nm/��4u����]��Y�IO��B�Oe���I�f�{ZS��#�,Y)�0�����N]TB)�����6�ꮤ�C�gx� �6�c�"pf�����M����52S�ʨb�͂9�,l9º�~��`�T,��o����}*8��dّ+�@���K$��de�����T�9�k�`�Ey|լ Xs/+WXQ/+k0/9VcVd9Xs/wDll0P/AKTT/wBVcVd9Xs/+WXQ/+k0/9VcVd9Xs/wDll0P/AKTT/wBV 72.00 0000660163 00000 n 0000441572 00000 n xmp.iid:B480E402F04EDF11986A9631617289C4 xmp.iid:2C25F82EE84EDF11986A9631617289C4 1rXNQ7pLf0zD/wBTQ3/cOGKu/TMP/U0N/wBw4Yq79Mw/9TQ3/cOGKu/TMP8A1NDf9w4Yq79Mw/8A Complexing protocol: siRNA is added to pre-diluted lipid in opti-MEM, mixed and complexed for 20 minutes. RihHfFWoJvI8kX+jvpTR3dswIQ25WW1EjK4PHZoxIWB7cq98VVzeeVopVuWubFZIAPTdpIv3QhX0 /UuS/wDcQT/mjFXfoJP+pcl/7iCf80Yq79BJ/wBS5L/3EE/5oxV36CT/AKlyX/uIJ/zRirv0En/U Horizon Team, Horizon/Perkin Elmer. Superior knockdown with Lipofectamine RNAiMAX Transfection Reagent compared to competing siRNA transfection reagents. After a short incubation period (30-90 min), cells are added rv0Z/wBqXVv+R/8A17xV36M/7Uurf8j/APr3irv0Z/2pdW/5H/8AXvFU9s/JmmXNrFPKb23eReTQ 0000555297 00000 n Found inside – Page 120Transfection was performed according to the manufacturer's protocol using 100 nM concentrations of each siRNA (siGENOME SMARTpool reagent Dharmacon, Lafayette, CO, USA) and 0.4 uL of DharmaFECT 1 siRNA transfection reagent (Dharmacon) ... Complexing time: 20 minutes. 2010-04-23T09:50:59-06:00 0000550778 00000 n /pjXGrXEw0y+uQ3D97BcLHG1EQfCpU0pSmah3SXfoV/+rNqf/SUn/NGKu/Qr/wDVm1P/AKSk/wCa /pAP/VTFW11G6dgg8xXlWIArYnv/AM9MVZdpGl6rYTPJf6o+ooy8VjaIR8TWvKodsVTVuh+WKrJZ Following a very simple protocol, the SMARTpool reagents are rehydrated in each well with a mixture of DCCR and DharmaFECT. DharmaFECT ® Duo is a lipid-based reagent that has been optimized specifically for co-transfecting plasmids and small RNAs, such as miRNAs and siRNAs. 0000563994 00000 n xmp.iid:D2FFF3F5C099DF11877BC7512233A314 This volume in the prestigious Methods in Enzymology series discusses methods currently used in preclinical and clinical gene therapy. 0000535111 00000 n Cell viability was determined 24 hours post-transfection using the CellTiter-Blue® Assay (Promega) and normalized to untreated cells. 0000446340 00000 n Be the first to comment on this protocol. 0000503669 00000 n 0000446448 00000 n Successful siRNA experiments require a transfection agent optimised for siRNA delivery, the efficiency with which different transfection agents deliver siRNA can vary. Vd9V0n/qXNR/4OT/AKp4q76rpP8A1Lmo/wDByf8AVPFV8UWlRc6eWr9vUUpVmc0r3X93scVWfVdJ v3afZ4oRt0zUO6Sv6rpP/Uuaj/wcn/VPFXfVdJ/6lzUf+Dk/6p4q76rpP/Uuaj/wcn/VPFXfVdJ/ 0000665989 00000 n 0000020466 00000 n ������c��Òp�|��H~���S��py�zn:x���F{b��I����w����F��2�����3]P%��m��1_Wh��� �+�K"��RI�=��;����v�>���hj���#�[r�j3����0w_������R�&"�M��1�٩��u�vK�;�%A1����o����ex�\���)b�@P��:�>�����,��a��v�p�~�cs��w>�ݴ�WWX�۳��4� C�o#�ẽ����B�,83�$a�rE���r�d_+�v��\Pw��(�}��:=��E����ǧ]�9��¹���;����De�U����2!���/�- �7�m�3^_J��+�H~��r�;�y�w^=Wt7j$z�d�Cڽ�%��^F=����5�[�vѱ����CDvҶ DharmaFECT® has three other transfection reagents named DharmaFECT® 2, 3 and 4 which have been tested for siRNA transfection with a variety of cell lines, including the . try 1-2 siRNA concentrations (ideally 10 nM or below; but for some tough cells or targets might need to climb higher), and eg 3 . 0000591336 00000 n 0000614555 00000 n Adobe PDF Library 9.0 68+9v+qmKu/wVff9X68+9v8AqpiqraeUby2uobltau5lhkSQxsW4uFIbif3h2NMVZPirsVSXzn/y Volume 1 provides a panel of 16 in vitro measures of chemosensitivity in adherent and non-adherent cells for single agents and combinations of agents. l+nFUy1e61y40hbiGOZp7e5kKQQRXFsb2NYZeMZ4lpberts7bclFdmxVDXnmTzMs0BstNZ5JFnWT 0000010673 00000 n 0000573297 00000 n 0000526177 00000 n For plasmid transfection, cells were transfected with 50 ng of pCMV6-RSU1 (Origene Technologies, Rockville, MD, USA) using DharmaFECT 1 Transfection Reagent (Thermo Fisher Scientific) for 12 h according to manufacturer's protocol. The protocol is similar to the suggested siRNA transfection protocol. This book analyzes all aspects of metronomic chemotherapy, a new approach involving low-dose, long-term, and frequently administered therapy that has preclinical and clinical activity in various tumors. 5,000 10,000 15,000 . 0000628583 00000 n Adobe InDesign 6.0 In case a particular cell line shows low transfection using DharmaFECT 4, try other transfection reagents such as DharmaFECT 1, 2, and 3 (Dharmacon, catalog nos T-2001, T-2002, and T-2003) or Lipofectamine RNAiMAX and Lipofectamine 2000 (Life Technologies, catalog nos 13778-075 and 11668-019), and TransIT-TKO siRNA Transfection Reagent (Mirus . 0000440693 00000 n 0000600880 00000 n Key features: DharmaFECT 1, 2, 3, and 4 : Multiple formulations for successful delivery of siRNA to any cell line. Mfr. 0000016399 00000 n GDamwL+pG85K3lybZuMcMiMWVVqKVqdsVSy58xeaYLfUboyaeo07Rk1gwiCWTk0y3jLCJ1u1Uqn1 0000530296 00000 n Version U1.1 extra to account for pipetting errors and evaporation. This book illuminates what we know, how we know and what remains to be discovered in this regard. The book explore and identify not only the key compounds of cell signaling pathway but also the interaction between them, the cross-talk. 0000447576 00000 n 0000021598 00000 n Scrambled small interfering RNA (siRNA) (D-001810-02) or S1PR2 siRNA (J-005208-06) were purchased from Dharmacon (Thermo Fisher Scientific, Lafayette, CO) and transfected into primary hPASMCs using DharmaFECT 1 transfection reagent (Thermo Fisher Scientific) according to the manufacturer's protocol. 0000021402 00000 n Catalog ID. 0000671203 00000 n 0000440518 00000 n Found inside – Page 291... FBS ( see Protocol 1 ) DharmaFECT 3 transfection reagent ( Dharmacon ) DharmaFECT cell - culture reagent ( Dharmacon ) Cell - cycle siARRAY RTF siRNA 96 - well imaging plates ( GE Healthcare / Cell Proliferation Fluorescence Assay ... nga/CNlcEgs5XxrhCpEthfjSxCLehafmp9SHiwKEbH1aYb3RSnJpuovHahLev7sxikkO7erM9F/e 0.2 ml; Sufficient for 50-100 transfections (per well in a 6-well plate) Enables highly efficient DNA transfection in a variety of cell lines including CHO-K1, COS, LNCaP, NIH/3T3, 293, T24, C2C12, SF-9, primary human keratinocytes, primary aortic smooth muscle, primary rabbit myoblasts . 0000598461 00000 n O/5FR/8ANGKp9Zy2k1rFJYlDbcQIvTFE4r8ICgdhTFXXkbTWc8KcOUkbqvqjlHUqQOa918RirFP8 Vp/1aNP/AO4k3/VbFV62FukbxDSNO4yU5V1Ak/CajixlJX6MVWfoq0/6tGn/APcSb/qtirv0Vaf9 APq/ab/0mQf9VMVd/jPyf/1ftN/6TIP+qmKu/wAZ+T/+r9pv/SZB/wBVMVd/jPyf/wBX7Tf+kyD/ 0000620353 00000 n 0000016646 00000 n 0000023334 00000 n 0000520518 00000 n 0000009336 00000 n 2010-04-23T09:50:52-06:00 53�,��ԅ4��X����W��j����uL��Qs�c�P]P�`MP���R�m[���Q�V Gm9�#�sC��-�y��Q:�O������\�{��`�0:Q{DuQ�$��V��1���*T #1��a��Lp����,t�^�xo�3�Ʃz��7:�����"��lȡ*59t+�@� ��Lt/昰�L�u�។�EI�#�([7.1a���(g�+H����S���qZ���qxf �4���S~����#{)I��7Frg�!a#�.M�R���!��K�@��11X��+ꋜ����M.H��i �� Y���`p��&�0C�A+. dk9tJawyi2MboslhM5lIpwkWOR6mtGP939nFVfX4/O9/p5sbKzuXiu7K4juRcSWKuJZYroRgGF0o

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