Our blogs and videos choice: dna sequencing gel

Engineering Mini project: DNA COMPUTERS

After seven days of intensive laboratory work, Adleman's test-tube contained the answer to the problem, subsequently visible as a series of dark bands on a DNA sequencing gel. On the face of it, it might hardly seem worth the bother, especially as Adleman already knew the answer before he started the experiment. But this was much more than a curious laboratory stunt. During the initial 'linking-up' stage of the process, Adleman's test-tube computer effectively performed an astonishing 10^14 calculations. And it did so with the consumption of only a tiny amount of energy, and in a tiny [more...]

Date: 2010-10-12 10:10:00

Blog posts (43) | Videos (26)

Can I use sheared genomic DNA as starting material of bacterial ...

My question is whether initially sheared genomic dna can be safely used for genome sequencing and assembly by 454 technology. By 'initially', I mean genomic DNA was sheared during extraction process. I attach gel image of my genomic dna samples (marker is lambda dna digested with EcoRI and HindIII). Although I know that dna fragmentation step is involved in standard genome sequencing by GS FLX titanium, I am concerned because I think it might be possible that shearing during extraction can affect randomness of standard fragmentation step and so on. Thanks in [more...]

Date: 2010-10-12 08:03:29

What are gel elctrophoresis and DNA microarray? How is each used ...

Question by neonpinkispink: What are gel elctrophoresis and DNA microarray? How is each used in the study of genetics? Best answer: Answer by brainybroad269 Gel electrophoresis is the use of a gel to separate pieces of DNA (or RNA, or protein) under an electric field. The simplest form separates pieces by size. More complex (2-dimensional) techniques can separate proteins by size and charge. A DNA microarray is a collection of short DNA sequences (“oligonucleotides”) immobilized on a piece of glass or other support (the “gene chip”). When you incubate the chip with your experimental [more...]

Date: 2010-09-25 04:09:26

Dr. Pandey “pulls off” new DNA sequencing method

The current method to sequence DNA is called gel-electrophoresis. This process is very costly and time consuming. However, Dr. Pandey and his research team have proposed a way to make DNA sequencing far easier and cheaper than the current method. For those of you scratching your heads and wondering what the purpose of DNA sequencing could possibly be, imagine in the futurew going to the doctor and getting customized medicine just for you based on your DNA. This medicine would do exactly what it needed to in order to fix your problem because it was made specifically for you and only you. [more...]

Date: 2010-09-23 17:36:11

Research Technician II (Durham) | PCR Jobs

Three years of molecular biology research and experience in the following is preferred: DNA/RNA extraction; DNA sequencing; PCR/ Real Time PCR; Taqman; gel electrophoresis; and tissue culture (propagation and storage). Computer proficiency in MS office programs, Photoshop, and basic statistical programs is also preferred. Qualified candidates should email a resume to the “reply to” email above. Please note “Tech II” in the subject line. Duke University is an Equal Opportunity/Affirmative Action Employer. Topics: Raleigh-Durham PCR jobs | No Comments [more...]

Date: 2010-10-11 18:43:40

Gel electrophoresis separates DNA fragments based on their…?

Question by Nicole Has-Ellison: Gel electrophoresis separates DNA fragments based on their…? A) recognition sequence B) length C) positive charge D) chemical bonds Best answer: Answer by BP B) length but others things like salt, whether the DNA is linear or a circle, whether the DNA is single or double stranded, whether the DNA is intact or has nicks in one or both of it’s strands all affect its speed of migration in a gel… What do you think? Answer [more...]

Date: 2010-10-09 04:10:59

ISPUB - Identification Of Lipase – Producing Psychrophilic Yeast ...

Lambda HindIII was used as DNA marker. After the electrophoresis, gel was stained with ethidium bromide and documented by the Gel Doc 2000 (Bio-Rad). Purification of PCR product was done by using QiAquick Gel Extraction Kit. Then, the purified PCR product obtained was sequenced and submitted for BLAST (Basic Local Alignment Search Tools) at NCBI (National Center for Biotechnological Information). Lipase assay by titrationActivity of lipase was assayed by titration using oil emulsion (polyvinyl alcohol: olive oil; 3:1) as substrate (Arima et al., 1972). The assay medium consists of 5ml oil [more...]

Date: 2010-08-25 00:51:13

O.N.L.Y 1 %: 10 Tips For Better DNA Gel Extraction Results

Nabi s.a.w bersabda, maksudnya: Jabir mengatakan bahawa Rasulullah s.a.w pernah bersabda, "Wahai manusia, aku telah meninggalkan untuk kalian 2 perkara. Jika kalian menggunakannya tidak akan tersesat selamanya iaitu Al-Quran dan Ahlul Bait keturunanku." (HR At-Tarmizi dan ia menghasankannya) What is it about gel extraction of DNA that makes it a pain? Maybe it’s poor product yields or maybe it’s because the process uses harsh chemicals (chaotropic salts, ethidium bromide, ethanol, heat) that will damage or denature DNA and potentially decrease cloning success. In this article I share some [more...]

Date: 2010-10-11 04:17:00

Summarize the way gel electrophoresis is used in DNA fingerprinting?

Question by princess100184: Summarize the way gel electrophoresis is used in DNA fingerprinting? please help Best answer: Answer by bio girl DNA is cut into segments using a specific restriction enzyme that cuts along specific sequences. the pieces are then run through electrophoresis and the different sized segments separate and can be compared to another sample. if both samples have the same segments cut by the same restriction enzyme they are the same person hope this helps Give your answer to this question [more...]

Date: 2010-09-23 04:15:54

Doe Contractor-grantee Workshop Sequencing Technologies

A bottleneck in gel-based systems is the manual gel-preparation step, which is both time consuming and a potential source of variability in DNA sequencing. Barry Karger (Northeastern University) discussed a fully automated, closed-end, [more...]

Date: 2010-07-05 19:44:31

iGEM UT-Tokyo(東大)実験ノート - 9/2 purification of DNA by ...

Experimenter Kariyazono, Nakatake, Zen Abstract purification of location sequence(digested by X & P on 9/2) by electrophoresis & extraction of bands from the gel Protocols 1.separating of DNAs through electrophoresis ←direction sample1 sample2 sample3 sample4 nothing(for 3 lanes) sample3′ sample4′ sample1′ 2.Extraction of DNAs Result;consentration of DNA sample1 5.3ng/ul sample2 7.8ng/ul sample3 6.9ng/ul sample4 7.3ng/ul sample1′ 7.8ng/ul sample3′ 5.8ng/ul sample4′ 6.8ng/ul flpe(PCR②) 3.6ng/ul stored in the [more...]

Date: 2010-09-02 04:27:41

DNA sequencing « Lenovo Laptop

Cognition of DNA sequences has gone indispensable for basic biological research, other inquiry legs utilizing DNA sequencing, and in numerous Gene Expression applied fields such as diagnostic, biotechnology, forensic biology and biological systematics. The advent of DNA sequencing has significantly accelerated biological inquiry and discovery. The rapid speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of the human genome, in the Human Genome Project. Related projects, frequently by scientific collaboration across continents, receive [more...]

Date: 2010-10-12 16:36:19

Bio Saga Blog: 5-Day Hands-on Workshop on Molecular Biotechnology ...

Recombinant DNA Technology: Cutting & pasting of DNA molecules (i.e. restriction digestion, ligation, DNA gel eectrophoresis, and gel extraction.) Bioinfo part: Bioinformatics of DNA database / SequenceAnalysis: Pairwise sequence alignment, Multiple sequence alignment, Pattern search.Tools: BLAST, CLUSTALW/X, CLC Bio Main Workbench Genetic Engineering: Transformation and plasmid purification, cloning and sub-cloning, amplification of DNA fragments by polymerase chain reaction (PCR). Bioinfo part: Designing of PCR primers In-silico cloning Searching for homologous and paralogous [more...]

Date: 2010-09-29 05:41:00

Sequenziamento del DNA- Prima generazione (metodo Sanger) « my GenomiX

Così, ecco la prima parte della mia serie sul DNA sequencing: oggi tratterò la prima generazione, vale a dire il metodo Sanger. Innanzitutto facciamo un piccolo ripasso sul DNA. Il DNA (o acido deossiribonucleico) è una molecola che ha l’aspetto di una doppia elica formata da due catene di nucleotidi. I nucleotidi sono piccole molecole costituite da un gruppo fosfato, uno zucchero (deossiribosio) e una base azotata; mentre le prime due componenti sono sempre uguali e costituiscono l’ossatura della doppia elica, le basi azotate esistono in quattro “versioni” differenti. Esse si [more...]

Date: 2010-09-08 08:31:06

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T4 Polynucleotide Kinase 500 u EK0031 450,000 T4 DNA Ligase (With PEG) 200 u EL0335 450,000 T4 DNA Ligase 200 u EL0015 450,000 T4 DNA Polymerase 100 u EP0061 440,000 Taq DNA Polymerase (native) 100 u EP0281 470,000 Taq DNA Polymerase (native) 500 u EP0282 1,880,000 Taq DNA Polymerase (recombinant) 100 u TA7505C 50,000 Taq DNA Polymerase (recombinant) 5,000 TA7507C 1.200,000 Taq DNA Polymerase (recombinant) 500 u TA7506C 160,000 TaqI 3,000 u RE7717C 230,000 TBE Buffer, 5X 50 ml MR7725C 20,000 Total Protein (Automatic and manual) 300Tests BK8145C 25,000 Triglycerides (Automatic and manual) New [more...]

Date: 2005-10-25 10:14:00

Thrombosis Journal | Full text | Expression of sterol regulatory ...

DNA was extracted from frozen cardiac muscle samples. The SCAP 2386A>G genotyping was based on PCR amplification, restriction enzyme analysis and DNA electrophoresis. The DNA samples were amplified by PCR, using the primers 5'-TTGTGCTGCGCGGCCACCTCA-3' and 5'-AGGAGGAAAGGGCAGCCGCAC-3'. PCR was performed in a volume of 50 μl. Cycle conditions were 94°C for 4 min, then 28 cycles of 94°C for 1 min, 64°C for 1 min and 72°C for 1 min, with a final extension step of 5 min at 72°C in a PTC-225 thermal cycler (MJ research, Massachusetts, USA). 10% DMSO was included in PCR reaction. The [more...]

Date: 2009-02-18 08:00:00

PLoS ONE: Analysis of Short Tandem Repeats by Parallel DNA Threading

Abstract TopThe majority of studies employing short tandem repeats (STRs) require investigation of several of these genetic markers. As such, we demonstrate the feasibility of the trinucleotide threading (TnT) approach for scalable analysis of STRs. The TnT method represents a parallel amplification alternative that addresses the obstacles associated with multiplex PCR. In this study, analysis of the STR fragments was performed with capillary gel electrophoresis; however, it should be possible to combine our approach with the massive 454 sequencing platform to considerably increase the [more...]

Date: 2009-11-13 08:00:00

Evaluation of Five Methods for Total DNA Extraction from Western ...

DNA extraction is a routine step in many insect molecular studies. A variety of methods have been used to isolate DNA molecules from insects, and many commercial kits are available. Extraction methods need to be evaluated for their efficiency, cost, and side effects such as DNA degradation during extraction. Methodology/Principal Findings From individual western corn rootworm beetles, Diabrotica virgifera virgifera, DNA extractions by the SDS method, CTAB method, DNAzol® reagent, Puregene® solutions and DNeasy® column were compared in terms of DNA quantity and quality, cost of materials, [more...]

Date: 2010-08-13 07:00:00

FinchTalk: From Reads to Data Sets, Why Next Gen is not like ...

The first widely-used sequencing systems were based on the “Sanger” method. DNA was synthesized in the presence of chain terminating radioactive dideoxy-nucleotides (1). Mixtures of DNA fragments were separated by size using gel electrophoresis and the bases were identified and entered into a computer through manual techniques. Automated DNA sequencing instruments arrived later. These instruments made DNA sequencing thousands of times more efficient by detecting fluorescently labeled fragments and sending the information directly to a computer (2). For the first time, it became possible [more...]

Date: 2009-01-06 19:16:00

MELANIE VIEWER 7.0 2D GEL ANALYSIS free download and reviews

Vector NTI Viewer 4.0.1 free download and reviews Size:520K Language: English Directory: Sequence Alignment and Analysis Requirements:Windows,MacOSX... MB DNA Analysis 6.84 free download and reviews Size:2984K Language: English [more...]

Date: 2010-10-01 11:25:03

Proteins – The common denominator of all diets

Complete DNA Sequencing & Synthesis Services,DNA Sequencing Analysis System. The human body is constantly changing with the environment. flow of matter and molecules in and out, casting in their complexity. Although the body lends them structure, it is the view – the food – who will decide his body. To control what goes in a diet is to decide what stays inside. Dietary decisions reflect an awareness of the metabolism and the nutrients needed to change it. It is said that a variety of diets for each activity and illness. Post a macronutrientwhich is always necessary in large quantities, [more...]

Date: 2010-10-07 15:50:25

ReportsandReports – DNA Sequencing in Drug Discovery: Market ...

Dallas, TX — (SBWIRE) — 10/04/2010 — The advent of DNA sequencing technology has had a vast impact on the field of drug development. It has played a key role in the success of the Human Genome Project which has made it an essential tool for genetic analysis. Advanced technological developments in microarrays, bioinformatics and similar tools have increased the scope of applications and services. This report covers regional markets, products and services while evaluating them over the next five years, and profiles almost 40 companies involved in the DNA sequencing market. Key features of [more...]

Date: 2010-10-05 21:38:18

Because no one will win the debate on human evolution!

All persons have the same 100% human genes and DNA sequence-based links to an African matriarch. I refer readers to 11th January 1988 cover story in Newsweek that the DNA of humans navigate on the River. This article has been shown that evolutionists have to search, but we are afraidWrite: All people are a unique rainbow family combined. We have the saliva, the same blood type, the same shit and the same disgusting habits: we are not many, but One Race A dysfunctional family! The fact that some pre-human species changed, does not mean that all people have done things before the same thing? [more...]

Date: 2010-10-09 17:50:37

CIENCIASMEDICASNEWS: Recurrent Granulibacter bethesdensis ...

DNA was isolated from human tissue by using the Maxwell 16 Tissue DNA Purification Kit (Promega, Madison, WI, USA) according to the manufacturer’s protocol. DNA concentrations were measured by using a UV spectrophotometer (NanoDrop, Wilmington, DE, USA). The 16S rRNA and methanol dehydrogenase subunit 1 (GeneID YP_744165.1) genes of G. bethesdensis were analyzed by using a PCR and primer sequences 16S-forward: 5'-TCGGGTGGGCACTCTAAAGG-3', 16S-reverse: 5'-GCATCACTGCCTAGCTTCCC-3', MDH-forward: 5'-CCGCAATACGGTCAATTCG-3', and MDH-reverse: 5'-GCCGATCTTCCAGGTTTCTTC-3'. Each reaction mixture [more...]

Date: 2010-09-02 23:42:00

Applied Biosystems 3500 Series Genetic Analyzers for DNA ...

The 3500 Series Genetic Analyzers set a new standard in capillary electrophoresis, with increased throughput, faster run times, and an innovative consumables system approach. To learn more, please visit: [more...]

Date: 2010-08-15 17:53:40

DNA Sequencing: New directions, and potential implications to ...

DNA Sequencing: New directions, and potential implications to healthcare Wired News (blog) However, much lower cost and more efficient methods for DNA sequencing will be required to make it feasible for routine healthcare practice and [more...]

Date: 2010-10-07 21:34:01

Market Manager, DNA Sequencing Applications

The Market Manager for DNA Sequencing Applications will be responsible for managing the DNA applications-based marketing activities necessary for the continued development and commercial support of the Illumina Sequencing business Job [more...]

Date: 2010-10-06 23:20:21

DNA Day at the St. Louis Science Center

Complete DNA Sequencing & Synthesis Services,DNA Sequencing Analysis [more...]

Date: 2010-10-09 05:00:09

Lab Tech / Research Associate (Torrey Mesa) | PCR Jobs

Candidate must be familiar with basic molecular biology techniques such as DNA purification, DNA gel electrophoresis, restriction digestion, PCR including primer design, plasmid construction, DNA sequence analysis software and bacterial transformation. Candidate will also assist with sample receipt and tracking, maintain inventory and equipment, and perform general lab maintenance and cleaning to include handling of biological waste. Education/ Skills/Attributes Required: · Must be a US Citizen. · BS degree in biology/biochemistry with 0-3 years industrial experience. · Experience with [more...]

Date: 2010-10-12 19:15:31


SSR assays require a minimal amount of genomic DNA. Most of the crop improvement programs are confined to evaluation and selection of naturally occurring clonal variations. In vitro culture techniques provide an alternative means of plant propagation and an important tool for crop improvement. The in vitro cultures are also of low risk for genetic variation since it is more resistant to genetic changes while occurrence of cell division in vitro condition. Gel electrophoresis is an important molecular biology tool which enables us to study DNA. It can be used to determine the sequence of [more...]

Date: 2010-09-08 04:07:38

Gene Sequencing: DNA analysis

Not everyone can properly and correctly analyze the DNA. Apartyears of study devoted to knowledge of DNA, DNA analysts to go through years of experience and practice in laboratories and practice of DNA testing and analysis in order to analyze the DNA and how it works. The training of DNA analysts understanding of sophisticated tools and machines developed for DNA analysis. proper management and use of these devices would make them more efficient in the pursuit of DNA to be analyzed correctly. Most of the DNA consists of 23Pairs of chromosomes, mitochondrial DNA inherited from the mother, and [more...]

Date: 2010-09-22 05:00:00

Genomics: The next generation | Wellcome Trust

The advent of the Human Genome Project and the development of faster, cheaper DNA sequencing revolutionised medical and biological research - and the technology is still evolving. Mun-Keat Looi looks at the platforms pushing us closer toward the ‘$1000 genome’. At the turn of the millennium, genomics captured the public's imagination with the publication of the first draft of the human genome and the insights it offered into our biology. When the Human Genome Project started in 1991, DNA sequencing entailed laborious radiation-based methods, with researchers manually loading [more...]

Date: 2009-07-28 11:00:00


GENOMIC CHARACTRIZATION OF AEROMONAS HYDROPHILA ISOLATES USING RAPD-PCR TECHNIQUE by Mitali Dhiman and S. S. Mishra Central Inland Fisheries Resear Aeromonas hydrophila are  involved in various disease problems in humans and aquatic  animals and are known to be  phenotypically, serologically and genetically quite diverse. Development and use of a sensitive and specific  diagnostic test is warranted for detection and characterization of this pathogen.In the present study,  Fish  and water samples from river Hooghly were streaked onto Aeromonas selective growth medium (Rimler-shotts [more...]

Date: 2010-10-08 20:08:05

JoVE: Electroeluting DNA Fragments (Video Protocol)

Purified DNA fragments are used for different purposes in Molecular Biology and they can be prepared by several procedures. Most of them require a previous electrophoresis of the DNA fragments in order to separate the band of interest. Then, this band is excised out from an agarose or acrylamide gel and purified by using either: binding and elution from glass or silica particles, DEAE-cellulose membranes, "crush and soak method", electroelution or very often expensive commercial purification kits. Thus, selecting a method will depend mostly of what is available in the laboratory. The [more...]

Date: 2010-09-05 22:00:00

csirhrdg.res.in | CSIR UGC NET 2011 Life Science Syllabus 2011 ...

Analysis of RNA, DNA and proteins by one and two dimensional gel electrophoresis, Isoelectric focusing gels. Molecular cloning of DNA or RNA fragments in bacterial and eukaryotic systems. Expression of recombinant proteins using bacterial, animal and plant vectors. Isolation of specific nucleic acid sequences Generation of genomic and cDNA libraries in plasmid, phage, cosmid, BAC and YAC vectors. In vitro mutagenesis and deletion techniques, gene knock out in bacterial and eukaryotic organisms. Protein sequencing methods, detection of post translation modification of proteins. DNA [more...]

Date: 2008-06-21 07:00:00

Gel electrophoresis separates DNA molecules on the basis of? - DNA ...

Question by Katie L: Gel electrophoresis separates DNA molecules on the basis of? Gel electrophoresis separates DNA molecules on the basis of A. the nucleotide sequence of their sticky ends. B. their nucleotide sequences. C. the amount of adenine they contain relative to the amount of thymine they contain. D. the amount of adenine they contain relative to the amount of guanine they contain. E. their lengths. Best answer: Answer by 6-Gun Annie Why are we doing your homework, of worse – your test – for you? I will be happy to explain anything you don’t understand, but help you [more...]

Date: 2010-10-11 04:07:47

DNA Gel Extraction Kit

The DNA Gel Extraction Kit is designed for the rapid purification of DNA fragments from agarose gels. The kit allows for the removal of agarose, enzymes, dNTPs, ethidium bromide, dyes and detergents from the DNA fragment. The recovered DNA is of a high quality, and is suitable for a number of downstream applications including restriction enzyme digestions, ligations, labeling, hybridizations and sequencing. The kit employs the use of a proprietary resin for DNA purification. This resin is an extremely inert and stable substance and is able to preferentially bind nucleic acids in the presence [more...]

Date: 2010-03-05 04:40:00

Forensic DNA was first used to convict criminals in 1985. Is this ...

Question by Morgan: Forensic DNA was first used to convict criminals in 1985. Is this the same thing as gel electrophoresis? I take AP bio but I obviously forget. Best answer: Answer by CryBabiesShouldntCry A short history from http://www.dna.gov/basics/analysishistory ‘DNA fingerprinting’ or DNA typing (profiling) as it is now known, was first described in 1985 by an English geneticist named Alec Jeffreys. Dr. Jeffreys found that certain regions of DNA contained DNA sequences that were repeated over and over again next to each other. He also discovered that the number of repeated [more...]

Date: 2010-10-03 04:23:14

Research Associate (San Diego) | PCR Jobs

"How To Answer Any Question An Interviewer Could Possibly Throw At You! ... " Click Here! Purpose of the Position:  Responsible for research and/or development of molecular diagnostic test kits, reagents and procedures in a team environment.  Performs experiments independently after receiving general direction from supervisor. Makes detailed observations, analyzes data and interprets results.  Prepares technical reports, summaries, protocols and quantitative analyses.  Maintains and develops skills in molecular biology and company technology through reading of internal reports [more...]

Date: 2010-10-01 19:07:24

DNA Art Blog » Blog Archive » PCR Process

We use primer sets that are optimally designed to span the specific genes you choose to have isolated for your canvas. Our technique focuses on the prevention of pre-PCR mis-priming and primer dimerization. Our primers are designed using software with algorithms that check for the potential of secondary structure formation and annealing of primers to itself or within primer pairs. We use hot start Taq Polymerase is designed to enhance the specificity, sensitivity, and yield of the target DNA sequence. Following PCR amplification and purification of the template DNA, we use restriction [more...]

Date: 2010-10-12 15:19:03

E. Coli Plasmid Vectors: Methods and Applications (Methods in ...

E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology) E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology) (archive) E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology (Clifton, N.J.), V. 235.) Table of Contents The Function and Organization of Plasmids Finbarr Hayes Choosing a Cloning Vector Andrew Preston Escherichia coli Host Strains Nicola Casali Chemical Transformation of E. coli W. Edward Swords Electroporation of E. coli Claire A. Woodall DNA Transfer by Bacterial Conjugation [more...]

Date: 2004-04-04 08:00:00

DNA sequencing » Blog Archive » DNA Sequencing Gel

Preparation of DNA Sequencing Gel 3. Assemble the gel rig on the sequencer, being careful not to get … 7. Run the gel for 45 minutes. E. Denature the sequencing reactions and prepare them for … DNA sequencing gel Hahn Lab Methods DNA sequencing gel. Acrylamide Urea Gel (100 ml) … if the top of the gel is wet with water and is wrapped tightly in Saran Wrap … How do we Sequence DNA? DNA Sequencing is at the center of the Human Genome Project, which promises to … sequencing gel: That’s exactly what we do to sequence DNA, then … Chapter 8 A. Recombinant DNA Technology PCR has [more...]

Date: 2010-09-28 04:49:52

Sandwalk: Nobel Laureate: Walter Gilbert

The Nobel Prize in Chemistry 1980. "for their contributions concerning the determination of base sequences in nucleic acids" Walter Gilbert (1932 - ) was awarded the Nobel Prize for developing a chemical method of sequencing DNA (with Allan Maxam). The method relied on chemical reactions that cleaved DNA at specific residues. By carrying out partial reactions where only one cleavage occurred in each DNA strand, it was possible to separate the cleavage products on an acrylamide gel and determine the position of each residue by the length of the fragment. The chemical sequencing strategy has [more...]

Date: 2008-12-10 14:35:00

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