The following dye-labeled terminator reaction chemistries have been designed to balance conservation of reagents with the resulting sequence product signal strength. When coupled with BACPAC Resource template preparation, N-free read lengths of >400nts can be expected. More emphasis can be placed on reagent conservation by further volume reduction or dilution; or on sequence product signal strength by increasing the reaction size. CycleSequenceConditions: (1) Initial denaturation step: 96oC 4min. (2) denaturation: 96oC 10sec. (3) annealing: 50oC 10sec. (4) extension: 60oC 4min. run steps 2-4 for 100 cycles. (5) hold: 4-10oC. Primers Useful For BAC and PAC Vectors: T7.29: 5’ gccgctaatacgactcactatagggagag 29mer T7: 5’ taatacgactcactataggg 20mer gSP6: 5’gttttttgcgatctgccgtttc 22mer 3Stock ABI Ready Reaction mixes are supplemented with 4mM MgCl2 4 Refers to ABI 1x (20ul) reaction 5 Annealing temp can be adjusted, based on Tm of primers used
|
→如果您认为本词条还有待完善,请 编辑词条
上一篇激光扫描共聚焦显微镜系统及其在细胞生物学中的应用 下一篇运动
词条内容仅供参考,如果您需要解决具体问题
(尤其在法律、医学等领域),建议您咨询相关领域专业人士。
0