33. Agarose Gel Electrophoresis
... • When the tracking dye reaches about one third to half of the length of the gel, it is the time to collect. • In general, 30 minutes are long enough to finish the electrophoresis process. Fig. 4 Migration of DNA fragments ...
... • When the tracking dye reaches about one third to half of the length of the gel, it is the time to collect. • In general, 30 minutes are long enough to finish the electrophoresis process. Fig. 4 Migration of DNA fragments ...
Document
... – mRNA from tissue of interest is isolated – Need to make double stranded DNA from mRNA: How? a. enzyme reverse transcriptase catalyzes synthesis of complementary single stranded DNA from mRNA i. Called complementary DNA (cDNA) because it is an exact copy of the mRNA b. mRNA is degraded either with ...
... – mRNA from tissue of interest is isolated – Need to make double stranded DNA from mRNA: How? a. enzyme reverse transcriptase catalyzes synthesis of complementary single stranded DNA from mRNA i. Called complementary DNA (cDNA) because it is an exact copy of the mRNA b. mRNA is degraded either with ...
Normalization between a pair of arrays
... This arises from the fact that while analyzing microarray data we conduct thousands of tests in parallel. Let we select 10000 genes with a significant level α=0.05 i.e a false positive rate of 5% This means we expect that 500 individual tests are false which is not at logical ...
... This arises from the fact that while analyzing microarray data we conduct thousands of tests in parallel. Let we select 10000 genes with a significant level α=0.05 i.e a false positive rate of 5% This means we expect that 500 individual tests are false which is not at logical ...
Lecture 9
... This arises from the fact that while analyzing microarray data we conduct thousands of tests in parallel. Let we select 10000 genes with a significant level α=0.05 i.e a false positive rate of 5% This means we expect that 500 individual tests are false which is not at logical ...
... This arises from the fact that while analyzing microarray data we conduct thousands of tests in parallel. Let we select 10000 genes with a significant level α=0.05 i.e a false positive rate of 5% This means we expect that 500 individual tests are false which is not at logical ...
UNIT 1: INTRODUCTION TO BIOLOGY
... the molecular process called meiosis kicks in. As you will see in the following sections below, sexual reproduction and the connected biological process meiosis assure the genetic material becomes shuffled and shuffled again during each generational cycle to purposely increase the genetic variabilit ...
... the molecular process called meiosis kicks in. As you will see in the following sections below, sexual reproduction and the connected biological process meiosis assure the genetic material becomes shuffled and shuffled again during each generational cycle to purposely increase the genetic variabilit ...
Cloning of genes from genomic DNA: Part 3
... Continuing from our isolation of genomic DNA and PCR amplification of either the evenskipped gene or the twist gene, we will now move on to the third step in the cloning procedure. We will use restriction enzymes to cleave off the ends of the PCR products. The oligonucleotide primers used in the PCR ...
... Continuing from our isolation of genomic DNA and PCR amplification of either the evenskipped gene or the twist gene, we will now move on to the third step in the cloning procedure. We will use restriction enzymes to cleave off the ends of the PCR products. The oligonucleotide primers used in the PCR ...
Bitter-Tasting Ability
... (RFLP) that can be separated on a 2% agarose gel. Each student scores his or her genotype, predicts their tasting ability, and ...
... (RFLP) that can be separated on a 2% agarose gel. Each student scores his or her genotype, predicts their tasting ability, and ...
E - Teacher Pages
... Abnormal numbers of sex chromosomes do not usually affect survival Sex chromosome abnormalities tend to be less severe as a result of – Small size of the Y chromosome – X-chromosome inactivation – In each cell of a human female, one of the two X chromosomes becomes tightly coiled and inactive – ...
... Abnormal numbers of sex chromosomes do not usually affect survival Sex chromosome abnormalities tend to be less severe as a result of – Small size of the Y chromosome – X-chromosome inactivation – In each cell of a human female, one of the two X chromosomes becomes tightly coiled and inactive – ...
on January 24, 2017 Downloaded from
... pigs were injected intradermally with 0.1 ml aliquots of 1:100, 1:1000 and 1 : 10,000 dilutions of antipurinoyl serum. Five hours later they were challenged by the intravenous injection of 0.5 ml of antigen solution mixed with 0.5 ml of 1 per cent Evans blue dye. Thirty minutes after this challenge, ...
... pigs were injected intradermally with 0.1 ml aliquots of 1:100, 1:1000 and 1 : 10,000 dilutions of antipurinoyl serum. Five hours later they were challenged by the intravenous injection of 0.5 ml of antigen solution mixed with 0.5 ml of 1 per cent Evans blue dye. Thirty minutes after this challenge, ...
Extrachromosomal DNA Transformationof Caenorhabditis elegans
... foreign DNA sequences in the transformed worm cells were measured by quantitative hybridization analysis. DNA preparations from transformed populations were spotted onto triplicate nitrocellulose filters. The filters were hybridized with 32P-labeled pBR322 DNA, bacteriophage DNA containing a single- ...
... foreign DNA sequences in the transformed worm cells were measured by quantitative hybridization analysis. DNA preparations from transformed populations were spotted onto triplicate nitrocellulose filters. The filters were hybridized with 32P-labeled pBR322 DNA, bacteriophage DNA containing a single- ...
Section 3: Prokaryotic Sample and Array Processing
... As starting material for the cDNA synthesis procedure, total RNA can be isolated by using standard procedures for bacterial RNA isolation or various commercial RNA isolation kits. For Pseudomonas aeruginosa and E. coli, we have successfully used the QIAGEN® RNeasy Mini Purification Kit. Caution shou ...
... As starting material for the cDNA synthesis procedure, total RNA can be isolated by using standard procedures for bacterial RNA isolation or various commercial RNA isolation kits. For Pseudomonas aeruginosa and E. coli, we have successfully used the QIAGEN® RNeasy Mini Purification Kit. Caution shou ...
chromosomes
... Staining procedures have been developed in the past two decades and these techniques help to study the karyotype in plants and animals. 1. Q banding: The Q bands are the fluorescent bands observed after quinacrine mustard staining and observation with UV light. The distal ends of each chromatid are ...
... Staining procedures have been developed in the past two decades and these techniques help to study the karyotype in plants and animals. 1. Q banding: The Q bands are the fluorescent bands observed after quinacrine mustard staining and observation with UV light. The distal ends of each chromatid are ...
Chapter 15 The Techniques of Molecular Genetics
... Circular, double-stranded circular DNA molecules present in bacteria. Range from 1 kb to over 200 kb. Replicate autonomously. Many carry antibiotic-resistance genes, which can be used as selectable markers. Many useful cloning vectors were derived from plasmid pBR322. © John Wiley & Sons, Inc. ...
... Circular, double-stranded circular DNA molecules present in bacteria. Range from 1 kb to over 200 kb. Replicate autonomously. Many carry antibiotic-resistance genes, which can be used as selectable markers. Many useful cloning vectors were derived from plasmid pBR322. © John Wiley & Sons, Inc. ...
Bacterial Genetics
... d. When you talk about bacteria that DNA or RNA can be isolated for example here is a gel agarose where we separate for example isolate the plasmid DNA. Here is a molecular standard here is a plasmid DNA that has been isolated and again it is an agarose gel i. Have the Agarose gel and you stain with ...
... d. When you talk about bacteria that DNA or RNA can be isolated for example here is a gel agarose where we separate for example isolate the plasmid DNA. Here is a molecular standard here is a plasmid DNA that has been isolated and again it is an agarose gel i. Have the Agarose gel and you stain with ...
Comparative genomic hybridization
Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells. The aim of this technique is to quickly and efficiently compare two genomic DNA samples arising from two sources, which are most often closely related, because it is suspected that they contain differences in terms of either gains or losses of either whole chromosomes or subchromosomal regions (a portion of a whole chromosome). This technique was originally developed for the evaluation of the differences between the chromosomal complements of solid tumor and normal tissue, and has an improved resoIution of 5-10 megabases compared to the more traditional cytogenetic analysis techniques of giemsa banding and fluorescence in situ hybridization (FISH) which are limited by the resolution of the microscope utilized.This is achieved through the use of competitive fluorescence in situ hybridization. In short, this involves the isolation of DNA from the two sources to be compared, most commonly a test and reference source, independent labelling of each DNA sample with a different fluorophores (fluorescent molecules) of different colours (usually red and green), denaturation of the DNA so that it is single stranded, and the hybridization of the two resultant samples in a 1:1 ratio to a normal metaphase spread of chromosomes, to which the labelled DNA samples will bind at their locus of origin. Using a fluorescence microscope and computer software, the differentially coloured fluorescent signals are then compared along the length of each chromosome for identification of chromosomal differences between the two sources. A higher intensity of the test sample colour in a specific region of a chromosome indicates the gain of material of that region in the corresponding source sample, while a higher intensity of the reference sample colour indicates the loss of material in the test sample in that specific region. A neutral colour (yellow when the fluorophore labels are red and green) indicates no difference between the two samples in that location.CGH is only able to detect unbalanced chromosomal abnormalities. This is because balanced chromosomal abnormalities such as reciprocal translocations, inversions or ring chromosomes do not affect copy number, which is what is detected by CGH technologies. CGH does, however, allow for the exploration of all 46 human chromosomes in single test and the discovery of deletions and duplications, even on the microscopic scale which may lead to the identification of candidate genes to be further explored by other cytological techniques.Through the use of DNA microarrays in conjunction with CGH techniques, the more specific form of array CGH (aCGH) has been developed, allowing for a locus-by-locus measure of CNV with increased resolution as low as 100 kilobases. This improved technique allows for the aetiology of known and unknown conditions to be discovered.