摘要:competent cells (DH5alpha at competency of 108 cfu/µl of pUC18). The transformation procedure it self is very similar to a standard CaCl2 transformation. Place 100 µl of TB buffer in a tub[阅读全文:]
摘要: 点击浏览该文件[阅读全文:]
摘要:PCR克隆主要有TA克隆法, 限制性酶切与连接法，杂交法和近期开发出来的特异性重组法等。但因为TA克隆法操作最简单，快速和高效的原因，成为Taq聚合酶PCR产物的最佳克隆方法。TA克隆法由Invitrogen发明，并拥有全球TA Clonin[阅读全文:]
摘要:Linker Ligation (with T4 ) DNA LigaseIn a microcentrifuge tube prepare a solution of blunt ended, dephosphorylated DNA (100-500ng) in TE buffer (5-7µl). Add 1-2µg of phosphorylated linkers[阅读全文:]
摘要:The overall sequence of events is:• Titer and plate out phage • Lift plaques onto filters and prepare them for screening • Make a probe • Hybridize the probe to the filters • [阅读全文:]
摘要:“多莉”的诞生，意味着人类可以利用动物的一个组织细胞 ，像翻录磁带或复印文件一样，大量生产出相同的生命体，这无疑是基因工程研究领域的一大突破。 人们剪下植物枝条，扦插到土里，不久就会发芽，长出新的植株[阅读全文:]
摘要:第一节 概 述 质粒具有稳定可靠和操作简便的优点。如果要克隆 较小的DNA 片段(＜10kb)且结构简单,质粒要比其它任何载体都要好。在质粒载体上进行克隆 ,从原理上说是很简单的，先用限制性内切酶切割质粒DNA 和目的DNA 片[阅读全文:]
摘要:TA CloningI. Initial mixture5 μl dd H2O1 μl PCR product1 μl ligation buffer2 μl TA vector (add second to last)1 μl ligase (add last)Incubate overnight at 15℃.II. Electroporation1. Chill[阅读全文:]
摘要:Instructions Modified for Simpson Lab1.PCR:Perform a standard PCR reaction using taq polymerase.(Do not use tfl as the PCR product must have a 3' adenine overhang that only taq generates.)Include an 7[阅读全文:]