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[核酸技术]
454测序法灵敏检测含量稀少突变株
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日期：2007-09-09 22:46:02
点击：99 评论：0
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| 454生命科学公司（454LifeSciences）、罗氏集团和一位耶鲁医学院研究院6月15日宣布，他们利用454公司的基因组测序系统，在一个早期经过临床治疗的样本中鉴定无法检测的稀有抗药性HIV突变。结果公布于巴巴多斯岛举行的第16国际HIV药物耐药会议，证实患者体内携带抗性突 |
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[核酸技术]
454测序新方法结合Sangon方法完善测序过程
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日期：2007-09-09 22:43:16
点击：227 评论：0
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| 自纽约2月13日的消息，美国能源部联合基因组研究所（DepartmentofEnergy'sJointGenomeInstitute，DOEJGI）的研究人员表示认为在微生物基因组测序方面表现出巨大的潜力的454LifeSciences公司的测序仪（见快速基因组测序时代到来）虽然并不能完全取代传统的Sanger测序方 |
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[核酸技术]
DNA序列测定技术
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日期：2007-03-26 08:23:21
点击：223 评论：0
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| 序列测定的技术和策略 Sanger双脱氧链终止法 Maxam－Gilbert DNA 化学降解法 测序策略 目前应用的两种快速序列测定技术是Sanger等（1977）提出的酶法及Maxam和Gilbert(1977)提出的化学降解法。虽然其原理大相径庭，但这两种方法都是同样生成互相独立的若干组带放射性标 |
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[核酸技术]
PREPARATION OF PLASMID DNA FOR SEQUENCING
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日期：2007-03-26 08:21:00
点击：74 评论：0
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| The following protocol is based on our modifications of R. Kraft, J. Tardiff, K. S. Krauter, and L. A. Leinwand. Biotechn . 6(6):544-545, 1988. Inoculate 2-5 ml of L broth containing the appropriate antibiotic from a single bacterial colony. Incubate |
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[核酸技术]
Single Strand DNA Prep. for Sequencing
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日期：2007-03-26 08:19:46
点击：79 评论：0
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| This protocol works well from a 5 ml starting culture or a 1 ml starting culture (adjust volume accordingly). Solutions 2X YT Media 16 g tryptone 10 g yeast extract 5 g NaCl 1 ml 1N NaOH up to 1 liter with Q Ampicillin Stock (1000X) 0.15 g ampicilli |
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[核酸技术]
PREPARATION OF PLASMID DNA FOR SEQUENCING
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日期：2007-03-26 08:16:38
点击：65 评论：0
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| The following protocol is based on our modifications of R. Kraft, J. Tardiff, K. S. Krauter, and L. A. Leinwand. Biotechn . 6(6):544-545, 1988. Inoculate 2-5 ml of L broth containing the appropriate antibiotic from a single bacterial colony. Incubate |
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[核酸技术]
Plasmid Sequencing
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日期：2007-03-26 08:15:33
点击：75 评论：0
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| This protocol gives M13 quality sequence when used with the Quick DNA Plasmid Prep. protocol D.2. Solutions Denaturation Solution 2 M NaOH 200 m l 10N NaOH 2 mM EDTA 4 m l 0.5 M EDTA pH 8.0 up to 1 ml with Q store at room temperature Precipitation S |
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[核酸技术]
DNA SEQUENCING (SANGER)
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日期：2007-03-26 08:13:17
点击：80 评论：0
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| DNA SEQUENCING (SANGER) (USB Sequenase Version 2.0) 1. For double-stranded DNA templates: a. Denature template: 10ml DNA (5-10g or alkaline lysis mini prepDNA) 8ml ddH2O 2ml 2N NaOH -incubate 30' @ 37oC. b. Dry-ice precipitate: +10ml 4M NH4OAc 100ml |
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[核酸技术]
DNA Sequencing
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日期：2007-03-26 08:10:08
点击：80 评论：0
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| Pouring the Gel Outline: Pouring this big thin 6% acrylamide gel ("Mother of all gels") is quite a challenge and probably the reason why smart whimps by them ready to use. Supplies Equipment: waterbath 37°C Erlenmeyer flask 250 mL with rubber stopp |
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[核酸技术]
Methods for DNA sequencing
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日期：2007-03-26 08:08:06
点击：87 评论：0
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| Methods for DNA sequencing A. Bst -catalyzed radiolabeled DNA sequencing Bst DNA polymerase-catalyzed radiolabeled two-step sequencing reactions (26) are modified from those presented earlier (25) by altering the absolute amounts and the relative de |
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[核酸技术]
放射性同位素标记的DNA序列测定分析
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日期：2007-03-26 08:03:54
点击：123 评论：0
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| 放射性同位素标记的 DNA 序列测定分析 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 聚合酶和 |
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[核酸技术]
Chemical Sequencing of DNA
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日期：2007-03-26 08:00:04
点击：54 评论：0
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| Chemical Sequencing of DNA 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) Biotechniques 2: 4-5. Steve H |
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[核酸技术]
DNA Sequencing
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日期：2007-03-26 07:57:07
点击：80 评论：0
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| DNA Sequencing The sequencing reactions described below work perfectly well if you are short of cash to buy sequencing kits. It is based on the Dideoxy sequencing method of Sanger et al., 1977. However, due to the number solutions that need to be ma |
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[核酸技术]
DNA SEQUENCING REACTIONS
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日期：2007-03-26 07:55:38
点击：67 评论：0
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| DNA SEQUENCING REACTIONS DNA priming reaction x ul DNA 2 ul Reaction buffer, minus DTT 1 ul primer (20 ng)10 ul totalHeat 90-100C 2 min.Cool room temp. 30 min. Enzyme mix 4 ul radioactive nucleotide (12 l) 2 ul reaction buffer ( 6 l) 4 ul water (12 |
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[核酸技术]
PREPARATION OF SEQUENCING GELS
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日期：2007-03-26 07:53:35
点击：68 评论：0
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| MATERIALS: 2-glass plates 1 sharks -tooth comb and spacers Whatman 3 mm paper 30 or 40% acrylamide-bis (19:1) 10X TBE urea 10% ammonium persulfate TEMED 60 cc. syringe Rain-ex Preparation of glass plates: Wash and rinse well the sequencing plates. S |
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