DNA重组

Single-stranded dye-terminator reactions required approximately 2 ug of phenol extracted M13-based template DNA. The DNA is denatured and the primer annealed by incubating DNA primer and buffer at 65d ...

Polyacrylamide gels for fluorescent DNA sequencing are prepared as described above except that the gel mix is filtered prior to polymerization. Optically-ground low fluorescence glass plates are caref ...

Sequencing double stranded DNA templates has become a common and efficient procedure (10) for rapidly obtaining sequence data while avoiding preparation of single stranded DNA. Double stranded templat ...

The following is a rapid and efficient method for sequencing cloned cDNAs based on PCR amplification (14) random shotgun cloning (1315) and automated fluorescent sequencing (16). This method was devel ...

用酚、氯仿法从工程菌中提取的质粒一般有三种构像，即超螺旋，线形和开环。（开环质粒是指双链环状的质粒DNA有部分解链，因此电泳速度最慢）三种构像的质粒在琼脂糖电泳的前后顺序是超螺旋线形开环，线形的质粒在中间，而开环的质粒在最后。判断质粒质粒好坏的一个指标就是超螺旋质粒在所以质粒中的含量。因为用质粒转染真核细胞时，超螺旋的质粒效率最高，所以要求质粒中超螺旋质粒的含量要在90％以上，这也是对作为核酸疫苗 ...

DNA序列测定分手工测序和自动测序，手工测序包括Sanger双脱氧链终止法和Maxam-Gilbert化学降解法。自动化测序实际上已成为当今DNA序列分析的主流。美国PE ABI公司已生产出373型、377型、310型、3700和3100型等DNA测序仪，其中310型是临床检测实验室中使用最多的一种型号。本实验介绍的是ABI PRISM 310型DNA测序仪的测序原理和操作规程。【原理】 ABI ...

Description:BiomolesSuperSpin Genome DNA Isolation Kitsare specifically designed to rapidly isolate super pure DNA from blood and tissue samples with utmost yields. The SuperSpin filter columns are ...

Objective:Preserve plasmid and genomic DNA.Storage:Open tube or remove seal from plateAdd 1-20μl of DNA to tube or well (from picograms up to 30μg)Dry samples completely at room temperatureOvernight ...

取腺病毒以m.o.i=5病毒量感染293细胞，待细胞病变达+++～++++时收获。沉淀细胞加2m1 TE重悬，分装四个1.5ml Eppendorf管，每管加蛋白酶K(20μg/μl)7μl和10% SDS 50μl37 C孵育1 h再加5mol/L NaCI 150μl置于4℃反应2ho 13000rpm离心30min，转移上清，用等体积酚、酚一氯仿/异戊醇、氯仿/异戊醇各抽提一次，抽提上清加入 ...

取腺病毒以m.o.i=5病毒量感染293细胞，待细胞病变达+++～++++时收获。沉淀细胞加2m1 TE重悬，分装四个1.5ml Eppendorf管，每管加蛋白酶K(20μg/μl)7μl和10% SDS 50μl37 C孵育1 h再加5mol/L NaCI 150μl置于4℃反应2ho 13000rpm离心30min，转移上清，用等体积酚、酚一氯仿/异戊醇、氯仿/异戊醇各抽提一次，抽提上清加入 ...

病毒感染宿主细胞后常常存在一些非整合的线形或环状双链DNA分子，通常称为赫特（Hirt）DNA。下面是一个试剂盒的步骤：GENMED 赫特（HIRT）法病毒DNA 纯化试剂盒1． 准备 个 孔细胞培养板的细胞（不同试剂盒的具体用量不一，说明书中有具体流程）2． 抽掉 孔细胞培养板里的 孔培养液3． 轻轻沿壁加入 毫升GENMED 清理液（Reagent A），清洗生长的细胞表面4． 小心抽掉清理液 ...

先进的基于焦磷酸测序（Pyrosequencing）的PSQ96 MA技术平台，能提供基于序列测定的SNP检测、等位基因频率分析和甲基化检测服务。另外，还提供短片段的测序服务（50-100 bp），适用于细菌和病毒分型等研究领域。 基于Pyrosequencing的SNP分析具有快速、高通量的特点，完成96个样品的SNP分析仅需10分钟。整个实验过程（包括PCR扩增、单链分离、Pyro ...

先进的基于焦磷酸测序（Pyrosequencing）的PSQ96 MA技术平台，能提供基于序列测定的SNP检测、等位基因频率分析和甲基化检测服务。另外，还提供短片段的测序服务（50-100 bp），适用于细菌和病毒分型等研究领域。 基于Pyrosequencing的SNP分析具有快速、高通量的特点，完成96个样品的SNP分析仅需10分钟。整个实验过程（包括PCR扩增、单链分离、Pyro ...

Always wear gloves and use barrier (filter) tips when performing this protocol. To prevent confusion and contamination never process more than 4 swabs at one time.Preparation:Before beginning put down ...

Always wear gloves and use barrier (filter) tips when performing this protocol. To prevent confusion and contamination never process more than 4 swabs at one time.Preparation:Before beginning put down ...

Genomic DNA isolation is performed according to the FBI protocol (27). After the blood samples (stored at -70degC in EDTA vacutainer tubes ) are thawed standard citrate buffer is added mixed and the t ...

Materials: see Solutions for Recipes TE buffer 10% (w/v) sodium dodecyl sulfate (SDS) 20 mg/ml proteinase K phenol\chloroform (50:50) isopropanol 70% ethanol 3M sodium acetate pH 5.2 Phase Lock ...

Materials: see Solutions for Recipes TE buffer 10% (w/v) sodium dodecyl sulfate (SDS) 20 mg/ml proteinase K phenol\chloroform (50:50) isopropanol 70% ethanol 3M sodium acetate pH 5.2 Phase Lock ...

A. Bst-catalyzed radiolabeled DNA sequencingBst DNA polymerase-catalyzed radiolabeled two-step sequencing reactions (26) are modified from those presented earlier (25) by altering the absolute amounts ...

直接测序法是建立在MSP基础上进一步深入研究CpG岛各个位点甲基化情况的方法。重亚硫酸盐使DNA中未发生甲基化的胞嘧啶脱氨基转变成尿嘧啶，而甲基化的胞嘧啶保持不变，行PCR扩增(引物设计时尽量避免有CpG，以免受甲基化因素的影响)所需片段，则尿嘧啶全部转化成胸腺嘧啶。最后，对PCR产物进行测序，并且与未经处理的序列比较，判断是否CpG位点发生甲基化。此方法一种可靠性及精确度很高的方法，能明确目的 ...