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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:]
摘要:原理：DNA分子水平上的多态性检测技术是进行基因组研究的基础。RFLP(Restriction Fragment Length Polymorphism,限制片段长度多态性)已被广泛用于基因组遗传图谱构建、基因定位以及生物进化和分类的研究。RFLP是根据不同[阅读全文:]
摘要:Adapted from Frommer et.al.* 1.Dilute DNA (up to 2 mg) into 50 ml with distilled H2O. 2.Add 5.5 ml of 2M NaOH. 3.Incubate at 37℃ for 10 minutes (to create single stranded DNA ). 4.Add 30 ml of 10 mM h[阅读全文:]