DNA SEQUENCING

描述(申请人提供)：综合性癌症中心内的多个实验室 中心(CCC)的研究项目需要自动化、高通量的DNA测序。因此，CCC成员必须能够使用以具有成本效益的、及时的方式提供DNA序列分析的设施。我们建议将现有DNA测序核心的服务扩展到癌症中心成员。该设施的具体目标是：1.为癌症中心成员提供DNA模板准备、引物设计和周期测序方法方面的技术支持和培训；2.通过使用平板凝胶和毛细管电泳法的专用应用生物系统DNA测序仪的供应和维护，提供最先进的自动化DNA测序能力；以及3.为序列编辑和分析提供技术支持。现有的DNA测序核心已经运行了15年，在过去5年中产生了3400万个碱基对的初级序列，为300多个用户提供服务。电流通过- 该设施的年产量超过18,000个测序反应。虽然最初是由UAB艾滋病研究中心建立并提供部分资金，但CCC成员约占该设施使用量的79%。在2003年使用这一设施的117名经过同行审查的独立资助的首席调查人员中，是CCC的成员。目前，该设施的资金主要来自按存储容量使用计费，每个测序反应需要10美元的费用才能满足运营成本。癌症中心所要求的支持将使我们能够降低每个反应的成本，从而使自动测序对CCC的研究人员来说更容易负担得起。

DESCRIPTION (provided by applicant): Numerous laboratories within the Comprehensive Cancer Center (CCC) require automated, high through-put DNA sequencing for their research programs. It is thus imperative that CCC members have access to a facility that provides DNA sequence analysis in a cost-effective, timely manner. We propose to expand the services of an existing DNA Sequencing Core to Cancer Center members. The specific aims of this facility are: 1. To provide technical support and training to Cancer Center members in DNA template preparation, primer design, and cycle sequencing methodologies; 2. To provide state-of-the-art automated DNA sequencing capabilities through the availability and maintenance of dedicated Applied Biosystems DNA Sequencers using both slab gel and capillary electrophoresis systems; and 3. To provide technical support for sequence editing and analysis. The existing DNA Sequencing Core has been in operation for 15 years generating over 34 million base pairs of primary sequence in the past 5 years serving over 300 individual users. The current through- put of the facility is over 18,000 sequencing reactions per year. Although originally established and partially funded by the UAB Center for AIDS Research, CCC members are accounting for roughly 79% of this facility's usage. Of 117 peer-reviewed, independently funded Principal Investigators who used this facility in 2003, 64 were CCC members. At the present time, the facility is mostly funded through chargebacks, necessitating a fee of $10 per sequencing reaction to meet operating costs. The requested support from the Cancer Center would allow us to reduce these costs per reaction, thus making automated sequencing more affordable for CCC investigators.