Plasmids Sample Clauses

Plasmids pBK-CMV expression vector. pMIGR1-eGFP Figure 2.1 plasmid maps of pBK-CMV and MIGR1
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Plasmids. The hDAGLa plasmid was constructed as described before.[38] Briefly, full length human cDNA of hDAGL-a was pur- chased from Biosource and cloned into mammalian expression vector pcDNA3.1, containing genes for ampicillin and neomycin re- sistance. A FLAG-linker was made from primers and cloned into the vector at the C-terminus of hDAGL-a. The plasmid was grown in XL-10 Z-competent cells and prepped (Maxi Prep, Qiagen). The sequences were confirmed by sequence analysis at the Leiden Genome Technology Centre. Transfection. HEK293T cells were grown to & 70 % confluency in 15 cm dishes. Prior to transfection, culture medium was refreshed (15 mL). A 3:1 (m:m) mixture of polyethyleneimine (PEI, 60 mg/well) and plasmid DNA (20 mg/well) was prepared in serum free culture medium and incubated for 10 min at rt. Transfection was per- formed by dropwise addition of the PEI/DNA mixture (2 mL/well) to the cells. 24 h post-transfection, the medium was refreshed and after 48 h cells were harvested. U2OS_ABHD6-GFP stable expression. Full-length human cDNA of ABHD6 (Source Bioscience) was cloned into mammalian expression vector pcDNA3.1, containing genes for ampicillin and neomycin re- sistance. The inserts were cloned in frame with a C-terminal FLAG- and GFP-tag. Plasmids were isolated from transformed XL-10 Z- competent cells (Maxi Prep kit: QiaGen) and sequenced at the Leiden Genome Technology Center. Sequences were analyzed and verified (CLC Main Workbench). One day prior to transfection U2OS cells were seeded to a 6 ▇▇▇▇▇ plates (& 0.5 million cells/well). Prior to transfection, culture medium was aspirated and a minimal amount of medium was added. A 3:1 (m m@1) mixture of polyethy- leneimine (PEI) (3.75 mg/well) and plasmid DNA (11.25 mg/well) was prepared in serum free culture medium and incubated (15 min, RT). Transfection was performed by dropwise addition of the PEI/ DNA mixture to the cells. After 24 h, transfection efficiency was de- termined by fluorescence microscopy and transfection medium was exchanged for selection medium containing 800 mg mL@1 G418. 48 h Post-transfection single cells were seeded to 96 ▇▇▇▇▇ plates in 100 mL selection medium. After 14 days, plates were in- spected for cell growth, clones were checked for ABHD6-GFP ex- pression by fluorescence microscopy (GFP channel). From here on, cells were grown in maintenance medium containing 400 mg mL@1 G418, and expanded in 12- and 6-▇▇▇▇▇ plates and 10 cm dishes. Inhibitor treatment. The medium was ...
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Plasmids. Table 2-4 Plasmids used in this study Name Cloning vector Insert Use Reference Table 2-5 Plasmids generated in this study Name Cloning vector Insert Generation of insert 5’ cloning site 3’ cloning site
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Plasmids. Unless otherwise agreed in writing by the Parties, Client will notify OXB in writing which of the following will apply to the Manufacture of a Vector:
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Plasmids. Unless otherwise agreed in writing by the Parties, Client will notify OXB in writing which of the following will apply to the Manufacture of a Vector: (a) Plasmids incorporating a GOI (genome/transfer Plasmid) suitable for Manufacturing Vector shall be procured and provided to OXB by Client at Client’s sole cost; (b) Plasmids incorporating a GOI (genome/transfer Plasmid) shall be procured by OXB on behalf of Client from an Approved Subcontractor and will be deemed Client Materials after Client’s payment therefor. [***]Pass-Through Costs relating to the manufacture of such Plasmids shall be charged by OXB to Client, such costs to include the direct cost of Plasmids, their transport and storage, and any import duties. In addition to such Pass-Through Costs, Client shall pay OXB a handling fee for procuring such Plasmids, which shall be [***] of such external costs and shall cover all OXB’s internal and other external costs associated with procuring such Plasmids (including stability testing performed by OXB). In such circumstances, OXB will notify Client in writing of the total costs for such Plasmids in advance of them being incurred. OXB will invoice Client following receipt by OXB of an applicable invoice from the Plasmid manufacturer. Title to such Plasmids shall pass to Client upon Client’s payment to OXB of the Pass-Through Costs (including the handling fee) for such Plasmids, and OXB shall inspect, store and handle such Plasmids in accordance with the provisions of this Agreement governing Client Materials. Notwithstanding any other provision of this Agreement, OXB accepts no liability for the performance of the Plasmid manufacturer under this sub-clause 3.8(b) however the Plasmids are sourced and in particular, in the event the Plasmid manufacturer fails to perform or delivers defective Plasmids to OXB, other than to the extent resulting from OXB’s gross negligence or wilful misconduct; or (c) Helper Plasmids, i.e. env, rev and gag-pol Plasmids, shall be procured by OXB from an Approved Subcontractor at OXB’s sole cost and will be deemed Components.
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Related to Plasmids

  • Hepatitis B Vaccine Where the Hospital identifies high risk areas where employees are exposed to Hepatitis B, the Hospital will provide, at no cost to the employees, a Hepatitis B vaccine.

  • Library Borrowing privileges available without charge. Upon retirement an employee shall be issued a permanent individual library card.

  • PROGENY Unmodified descendant from the MATERIAL, such as virus from virus, cell from cell, or organism from organism.

  • Influenza Vaccine Upon recommendation of the Medical Officer of Health, all employees shall be required, on an annual basis to be vaccinated and or to take antiviral medication for influenza. If the costs of such medication are not covered by some other sources, the Employer will pay the cost for such medication. If the employee fails to take the required medication, she may be placed on an unpaid leave of absence during any influenza outbreak in the home until such time as the employee has been cleared by the public health or the Employer to return to the work environment. The only exception to this would be employees for whom taking the medication will result in the employee being physically ill to the extent that she cannot attend work. Upon written direction from the employee’s physician of such medical condition in consultation with the Employer’s physician, (if requested), the employee will be permitted to access their sick bank, if any, during any outbreak period. If there is a dispute between the physicians, the employee will be placed on unpaid leave. If the employee gets sick as a reaction to the drug and applies for WSIB the Employer will not oppose the application. If an employee is pregnant and her physician believes the pregnancy could be in jeopardy as a result of the influenza inoculation and/or the antiviral medication she shall be eligible for sick leave in circumstances where she is not allowed to attend at work as a result of an outbreak. This clause shall be interpreted in a manner consistent with the Ontario Human Rights Code.

  • New Technology When new or updated technology is introduced into a workplace, it will be the responsibility of the employer to provide appropriate and, if necessary, ongoing training to the employees directly affected. Such training will include any health and safety implications or information that will enable employees to operate the equipment without discomfort and will help maintain their general well-being.