2×Taq Master Mix (E005)

目录 规格 价格
产品描述:
本制品是将PCR反应所需的Taq酶、dNTP混合物、MgCl2以及反应缓 冲液预先配制成2倍浓度的混合物。2×Taq Master Mix专为常规PCR 扩增反应优化,扩增长度可达8kb,能对4kb及其以下长度的片段进行高效地扩增。使用时只需在加入模板和引物并稀释到1倍浓度即可进 行PCR反应,大大地简化了操作过程,减少了PCR操作过程中的污染。
2×Taq Master Mix提供普通型/快速上样型两种形式供您选择。经测试,染料的加入不影响PCR反应,在PCR反应完成后可直接电泳,节省时间。
产品特点:
  • 高效:以λDNA为模板,扩增长度可达8kb,能高效扩增≤4kb片段(图例一)。
  • 灵敏:可从0.05ng人基因组DNA模板中扩增出特定基因片段(图例二)。
  • 稳定:反复冻融几十次,4oC放置30天,室温放置一周后,扩增性 能不受影响。
  • 快捷:PCR反应所必需试剂全集于一管之中,数分钟即可完成反应体系配制。
  • 便利:10μl, 25μl 或50μl体系全部适用。PCR反应后可直接电 泳(含染料形式)。
产品用途:
  • 常规PCR鉴定;
  • 小片段目标基因克隆;
  • 平端PCR产物加A。
使用建议:
使用本试剂扩增得到的PCR产物3´ 端有一突出"A"碱基,可直接克隆于T载体中。当扩增较长片段时,推荐使用2×Taq plus Master Mix, Taq plus 对于较长的DNA片段扩增具有更高的扩增效率及保真度。
应用实例:
Taq DNA Polymerase Amplification
图例一)使用普通(A组)及含染料(B组)的2×Taq Master Mix配制的50μl扩增体系中,以5ngλDNA为模板,对500bp~6.0KB片段的扩增结果。
泳道1:500bp; 泳道2:1kb; 泳道3:2kb; 泳道4:4kb; 泳道5:6kb;泳道M:1kb DNA LadderI
Taq DNA Polymerase Amplification
图例二)50μl扩增体系中,分别以50ng~0.05ng人基因组DNA为模板,可 对特定基因片段(380bp)进行很好的 扩增。
泳道1:50ng; 泳道2:5ng; 泳道3:0.5ng; 泳道4:0.05ng;泳道M:DNA Ladder 2000

产品包装(A包装):

组成体积
2×Taq Master Mix1ml×3支

常用反应体系(50μl)

组分体积
2×Master Mix*25μl
上游引物0.2-1.0μM(终浓度)
下游引物0.2-1.0μM(终浓度)
模板1-50ng(质粒)
10ng-1μg(基因组)
ddH2O至50μl
*Mg2+终浓度为2mM

常用PCR循环:

当扩增片段<3K:
94 C 1分钟30秒
30次循环94℃,20秒/57℃,20秒/72℃,根据产物长度调整, 60秒/kb
72℃5分钟
4℃保温
当扩增片段≥3K(推荐引物长度≥30bp):
94 C20秒
30次循环 94℃,5秒/68℃,根据产物长度调整, 60秒/kb
72℃5分钟
4℃保温
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For research use ONLY.