摘要:A simple, reversible way to a stop restriction reaction is by adding EDTA, which chelates Mg2+ , thereby preventing catalysis. If further manipulations of the digested DNA are to be performed, the res[阅读全文:]
摘要:For every 4 mls of culture, dissolve the BAC DNA pellet in 40 µl of water. for example: Usually each BAC is grown in 20 mls LB/CM total, then is dispensed into one Autogen tube (4 mls in each of[阅读全文:]
摘要:16 µl BigDye Terminator Ready Reaction Mix1 µg BAC DNA (determined from gel)10 pmol primerwater to 40 µlHeat tubes at 95℃ for 5 min., then perform 30 cycles of:95℃ x 30 sec55℃ x 10 s[阅读全文:]
摘要:MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc. syringeRain-exPreparation of glass plates:Wash an[阅读全文:]
摘要:测定DNA 的核苷酸序列是分析基因结构与功能关系的前提。从小片段重叠法到加减法、双脱氧链终止法、化学降解法、自动测序，DNA 测序技术发展很快。目前在实验室手工测序常用Sanger双脱氧链终止法。Sanger法就是使用DNA 聚合[阅读全文:]
摘要:本方法的核心部分是医科院基础所的生化脂蛋白组的吴刚老师所创，我在其基础上进行了一些改动，主要是DNA回收上采取了更简单的方法。（本方法最适用于双酶切制作片断并进行克隆 的情况。对于分步酶切制作片断，也可以使用本[阅读全文:]
摘要:This is a rapid method for chemical DNA sequencing which is commonly used as ladder for footprinting reactions or for sequencing of short DNA oligonucleotides.Reference: Bencini et al. (1984) Biotechn[阅读全文:]
摘要:This protocol is intended to make shotgun libraries from BACS that are to be fully sequenced and assembled. To minimize chimeric clones, an adaptor method is used. There are simpler protocols if just [阅读全文:]