生物化学技术

问：刚做克隆，遇见测序过程中 有3'端测序，5'端测序，到底是怎么回事？为什么有两个测序结果？我应该按那一个来看测序正确还是不正确？ 答：测序都是从5'端进行的，正向和反向测序是指对DNA的两条互补链分别测序，通常两个方向测序结果经校读后完全一致才能认为得到可靠结果。一点测序经验： 1、测序选择引物很重要，一般的克隆载体现在都有通用引物，如T7SP6M13-47 RVM ....但选择哪个引物测序 ...

Q:my group is involved in a national project to develop a genomic platform for cancer research. while Nimblegen has developed target enrichment chip for the Genome Sequencer 20 nothing similar has bee ...

Q:We recently completed a ChIP-seq run on the ABI SOLiD. We received the following data back for one of our samples. These data were from a quarter (1/4) of a slide. ------------------61993194 total b ...

Q:I've been getting mixed messages about the accuracy of the reads coming off the SOLiD platform. A 454 representative swore blind to me last week that only 25% of SOLiD reads are error-free (!) but t ...

Sequences for Solexa Library Preparations: Genomic DNA oligonucleotide sequences Adapters 1 5' P-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG 5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT PCR Primers 1 5' AATGATACGGCG ...

Here I present a brief overview of Solexa's sequencing-by-synthesis chemistry. The sample prep methods used differ slightly from that used in ABI's SOLiD system but the basic goals are the same: gener ...

The following derives largely from an email I sent to some "fellow travelers" before I heard of this web site. Consider 3 SOLiD runs we did all genomic DNA (fragment 35mers single flowcell ...

Applied Biosystems has just launched their instrument which supports their version of high-throughput sequencing chemistry termed “SOLiD™” (little “i” please). Acquired f ...

The Solexa sequencing platform comprises all hardware software procedures and kitted reagents for DNA sequencing at the whole genome or repetitive regional scale. a) Preparation of full diversity libr ...

DNA测序原理和方法 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 ...

在分子生物学研究中，DNA的序列分析是进一步研究和改造目的基因的基础。目前用于测序的技术主要有Sanger等（1977）发明的双脱氧链末端终止法和Maxam和 Gilbert（1977）发明的化学降解法。这二种方法在原理上差异很大，但都是根据核苷酸在某一固定的点开始，随机在某一个特定的碱基处终止，产生A，T，C，G四组不同长度的一系列核苷酸，然后在尿素变性的PAGE胶上电泳进行检测，从 ...

Introduction For over 25 years Applied Biosystems has been a pioneer in the field of genetic analysis by offering systems to address the expansion of genetic analysis applications and the evolving ne ...

ABI SOLiD sequencing is a form of DNA sequencing. DNA sequencing encompasses biochemical methods for determining the order of the nucleotide bases adenine guanine cytosine 5-methyl cytosine and thymin ...

ABI SOLiD sequencing is a form of DNA sequencing. DNA sequencing encompasses biochemical methods for determining the order of the nucleotide bases adenine guanine cytosine 5-methyl cytosine and thymin ...

technology: illumina sequencing technology This breakthrough platform is based on massively parallel sequencing of millions of fragments using our proprietary reversible terminator-based sequencing ch ...

One Fragment = One Bead = One ReadThe complete sequencing workflow of the Genome Sequencer FLX System comprises four main steps leading from purified DNA to analyzed results. These basic steps include ...

454 Life Sciences is a biotechnology company based in Branford Connecticut specializing in high-throughput DNA sequencing using a novel massively parallel sequencing-by-synthesis approach. 454 has exp ...

一、测序的基本原理： 测序的基本原理是Sanger末端终止法，在酶的作用下将原本是一条长达数百碱基的片断扩增成数百条片断，彼此间相差一个碱基。用电泳分离这些片断后，再通过测序仪将这些信号转变成峰图，从而最终形成客户所看到的测序序列和测序的峰图。 目前通用的扩增方式是通过PCR的方式进行的起源；其原理简述如下：PCR类似于DNA的天然复制过程，其特异性依赖于与靶序列两端互补的寡核苷酸引物。PCR由变 ...

DNA的复制需要：DNA聚合酶，单链DNA模板，带有3'-OH末端的单链寡核苷酸引物，4种dNTP（dATP、dGTP、dTTP和dCTP）。聚合酶用模板作指导，不断地将dNTP加到引物的3'-OH末端，使引物延伸，合成出新的互补DNA链。如果加入一种特殊核苷酸，双脱氧核苷三磷酸（ddNTP），因它在脱氧核糖的3’位置缺少一个羟基，故不能同后续的dNTP形成磷酸二酯键。如，存在ddCT ...

问：在实验室里做工艺，我用到2万转的告诉离心机进行微滤的前处理，不知道生产中的转速能达到这个吗？ 答：我们这里有13000转的。 答：管式离心机可达到1万转以上。 ...