1586/14737159. and immunoblotting (for protein; also known as Western blotting). Western Blotting Technique. Dot Blot (DB), Slot Blot (SB), and Colony Immunoblot are also techniques for detecting, analysing, and identifying proteins. 05% Tween-20, pH 7. Far–western blot analysis is an alternative method to analyze protein–protein interactions that entails probing proteins that have been separated by gel electrophoresis with tagged bait proteins, and then detecting the interacting proteins (prey) through multiple methods. Done after a gel electrophoresis, transferring the molecules from the gel onto the blotting membrane or adding the samples directly onto the membrane. S. 1. RNA blot hybridzation RNA blot hybridization is a technique that involves the detection of specific target within total RNA or mRNA that has been immobilized to a filter membrane. Mancha oscura en la posición de la membrana que contactó con una colonia que portaba la secuencia diana. Dot Blotting; SOUTHERN BLOTTING. Or • A northern blot is a method used to detect specific RNA molecules among a mixture of RNA and also be used to analyze a sample to measure. Zeder-Lutz et al. Dot blot technique is also called slot blot technique. g. The cells can be rapidly lysed and directly spotted on the nitrocellulose membrane. What is a Northern Blotting ? The northern blotting is a technique used in molecular biology research to study gene expression by detection of RNA. Dot blot hybridization is now the most commonly used procedure for testing of large numbers of samples. Northern Blotting By: Dr. We have developed a dot-blot assay specifically for use with a LiCor Odyssey CLx imager, which allows for sensitive detection of proteins in the infrared range. The sensitivity of the reverse dot-blot PCR was 92. 2, 2019. 1 Visualization by Transillumination 33 2. Your equipment, sample, antibody, and detection chemistries all can impact your results. The two most common methods for protein transfer are: Electrophoretic transfer — proteins separated in gels (for example, following polyacrylamide gel electrophoresis, or PAGE), are transferred onto membranes by electrophoretic transfer. D. • 10. dot blot technique synonyms, dot blot technique pronunciation, dot blot technique translation, English dictionary definition of dot blot. Southern. The reproducibility of the dot blot technique developed for this study was investigated by exposing serum of one healthy blood donor. The classical western blot has been used to monitor some proteins as putative biomarker in epilepsy (Tan et al. immunoblotting analysis immunoblotting and dot blotting immunoblotting applications immunoblotting assay immunoblotting definition immunoblotting experiment immunoblotting explained immunoblotting ib immunoblotting pdf immunoblotting ppt immunoblotting principle immunoblotting procedure immunoblotting protein. Lectut btn-202-ppt-l23. The technique was discovered by Edwin. 3. and molecular techniques in diagnostics is represented in Table1. The probe hybridizes to its complementary sequence and the hybridization product is detected with a. Western Blotting (also called immunoblotting) is a technique used for analysis of individual proteins in a protein mixture (e. ; 3. 2. BLOTTING • A molecular technique of transferring proteins, DNA or RNA, onto a carrier or membrane. Le Dot Blot est une technique simplifiée de Western Blot, qui est principalement utilisée pour la détection de protéines. 3. Dot-blot is generally a simple, fast and sensitive technique that enables to transfer a known amount of sample onto an inherent support, such as a Nylon membrane. materi Teknik pelacakan molekuler-genomik. This review is aimed to summarize some of the most commonly used techniques to determine DNA–protein interactions. In DB or SB the sample is applied directly to the membrane as dots or as slots using a vacuum apparatus. 2. pdf), Text File (. Once transferred to a membrane, they have to be. The blotted nucleic acids are then used as target in the hybridization experiment for their specific detection. Western blotting: Western blotting is an Immunoblotting technique which rely on the specificity of binding between a molecule of interest and a probe to allow detection of the molecule of interest in a mixture of many other similar molecules. Western blotting is an important procedure for the immunodetection of proteins, particularly proteins that are of low abundance. Immuno-northern blotting using antibodies against modified nucleosides. 3% comparing to. Briefly, DNA is. Blot 10 µl of 100 µg/ml of primary antibody onto membrane. A “sandwich-type” dot-blotting improves the usability of fluorescent detection technique To improve the user-friendliness and the specificity of the detection technique above, we devised a “sandwich-style” dot blot assay employing both anti-Ac and mAb-29, in which the bacterial cells were concentrated by the anti-Ac pre-coated on the. HISTORY 2The Oxford College of Science 3. 8K views• 15. Andri Setiawan; View full-text. Blotting techniques are commonly used analytical tools which facilitates identification of desired DNA/RNA/ Protein fragments. As a result subsequent blotting. In this method labelled antibody against particular protein is used identify the desired protein, so it is a specific test. 22. IMMUNOBLOTTING • Viral antigens are detected with a polyclonal or a MAb onto nitrocellulose paper. 4. g. Leana Kirei. The in-house reverse dot blot hybridization (RDBH) assay has been widely used in the spoligotyping technique for M. The technique is actually very similar to the Western blot, but dot blot, for reasons we’ll cover later, is a faster, cheaper, and easier technique. . This topic part has two sections: Content Tutorial and Animations. DNA-binding proteins control various cellular processes such as recombination, replication and transcription. Dot-blotting technique is particularly useful in obtaining quantitative data for the evaluation of gene expression. Receptor tyrosine kinases. 3. Edwin Southern (1975). There is also lot of blotting methods like nothern,southern,western blotting. Darker dots indicate more protein. The PCR-RDB assay consists of multiplex PCR amplifications of 10 fragments in the target sequence of. Western blot experiments typically include a small number of observations. coli strains, dot blot technique was performed. In case the blot still has a lot of background counts, wash the blot with wash solution IV at room temperature initially for 5 min and if required for another 10 min (see Note 29). Practical # 13 9-01-2019 Western Blotting Introduction: Blotting is the technique in which nucleic acids, proteins are immobilized onto a solid support generally nylon or nitrocellulosemembranes. Western blotting Ashfaq Ahmad. • Since, Nucleic acid hybridization is a process used to identify specific DNA sequences. The dot blot procedure is a well-validated technique, and its use is an alternative to avoiding the protein separation process although the equipment available. 6. Southwestern blotting is a technique used to study DNA-protein interactions. This technique is helpful in detecting the. The technique is a modified version of the Southern Blotting, which was discovered for the analysis of DNA sequences. Denature DNA 5. Blotting techniques ppt. A Southern blot (also called a Southern Transfer) is named after Ed Southern, its inventor. D. •Used to identify specific molecules in a complex mixture of related molecules. 4. Be able to explain the various types. Adapun tujuan dilakukan praktikum Dot Blot ini adalah untuk : 1. Recommended. WESTERN BLOTTING Western blotting, also known as Immunoblotting or Protein blotting, is a core technique in cell and molecular biology. A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. 5. 7. 45, 1. 6 Related techniques. Pulse Field Gel Electrophoresis Pankaj Gaonkar 21. • Blotting are techniques for transferring DNA, RNA and proteins onto a carrier so they can be separated, and often follows the use of a gel electrophoresis. 5 % when compared to direct DNA sequencing for detecting β-thalassemia mutations. The dot blot, or slot blot, is a technique which can be used for the detection of viral antigens from a sample (Li et al. 4. • Allow to identify a particular protein of interest. Blotting techniques Dr. 3. 5K• slides. Transfer to nitrocellulose paper (blotting) 6. 2. Southern who developed this procedure at Edinburgh University in the 1970s. Bar graphs are the least informative way of presenting continuous data as they conceal the spread of the data and sample size [15,32]. Western blotting involves the transfer of proteins that have been separated by gel electrophoresis onto Dot-blot hybridization provides a simple, easy, and fast way to quantify the relative concentration of ANF mRNA within cardiac tissue. Far–Western Blot Analysis. 9K views•30 slides. 4. Different blotting is used to detect different type of macromolecules such as southern blotting is used for DNA analysis, western blotting is for protein analysis, northern blotting is for RNA analysis and eastern for carbohydrate detection. The main steps in dot blot hybridization are: (1) a small amount of sap is extracted from the plant under test; (2) the viral nucleic acid is denatured by heating or, if it is DNA, by alkali treatment; (3) a spot of. 1. • Nucleic acid hybridization can be done in all. A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. We developed a PCR-reverse dot blot (RDB) assay for screening 20 hotspot mutations of GJB2, GJB3, SLC26A4, and MT-RNR1, which are common non-syndromic hearing loss (NSHL)–associated genes in the Chinese population. Take the membranes out of the plastic bins. Dot blot . The process of western blotting involves the transfer of proteins. FAR EASTERN BLOTTING 9. This technique is similar to the western blot technique. Western blotting • The Western blot is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. Il a été nommé d'après le Southern blot technique qui détecte pour l'ADN, inventé par Edwin M. Protein Blotting Methods. , Southern, dot blots, NPA Oligonucleotide End labelling • 3' end labeling of DNA is usually carried out using terminal transferase in template-independent manner to form a homopolymeric tail. This protocol provides the general workflow to simply entrap nucleic acids (in this case DNA) onto a Nylon membrane [ 1 – 3 ]. Southern Blotting A Southern blot is a method used in molecular biology for detection of a specific DNA sequence in DNA samples. This is termed slot or dot blotting and provides a convenient. This technique is based on the principle of separation of DNA fragments by gel electrophoresis and identified by labelled probe hybridization. The method is named after its inventor, the British. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. 6 GENE PROBE DERIVATION. they allow a higher degree of labeling to be achieved than end-labeling techniques. Autoradiograph showing the extent of sequence similarity between MSV-MNM and other MSV strains. Prasad Naidu 135. 5: Practical Applications of Monoclonal and. 6. Immunoblotting, also dubbed Western blotting, exploits antibody specificity to identify target proteins among a number of unrelated protein species (1,2). TYPES OF BLOTTING TECHNIQUES Blotting technique It is used to detect protein. A protein sample is subjected to electrophoresis on an SDS- polyacrylamide gel. However, while agaroses gel are typically. M andBlotting Techniques. The technique was developed by E. The proteins are then transferred to a membrane (typically nitrocellulose),. Neurologic Evaluation and Ancillary Tests. 1 Citations Abstract The techniques of Southern and Northern blotting have become widely known and are extensively used in the study of molecular genetics. The use of polyacrylamide gel electrophoresis is a prerequisite for western blotting in order to separate proteins prior to their identification. Blotting Techniques. Western blotting The western blot is a widely used analytical technique used to detect specific proteins in a biological sample. , 60 (Suppl 3) (2009), pp. IntroductionCommon techniques include: Southern blotting (DNA) , Northern blotting (RNA) , and immunoblotting (for protein; also known as Western blotting). Since dot-blot analysis with competitive hybridization provides. Develop the film the following day. G. 5. It represents a simplification of the western blot method, with the exception that the proteins to be detected are not first separated by electrophoresis. , 2011). SUBODH SHAH UTU. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concentrations of tissue samples. carrier or membrane. 3. Abstract and Figures. Blotting techniques includes southren,northern,western and dot blotting. For larger volumes or when making quantitative measurements, dot. In the next step, an alkaline phosphatase (AP) labelled. Northern blot is a technique based on the principle of blotting for the analysis of specific RNA in a complex mixture. 4. Moreover, the simplicity of conducting the northern analysis experimentally as well as its economic feasibility has globally propelled this technique as the method of. 2 ) DNA fragments are blotted onto a strip of nitrocellulose or a nylon membrane. In vitro techniques such as footprinting assays, electrophoretic mobility shift assay, southwestern blotting, yeast one-hybrid assay, phage display and proximity. Cell-free dot blot procedure. Western blotting techniques Southern blotting Northern blotting Western blotting Molecule detected DNA (ds) mRNA (ss) Protein Gel electrophoresis Agarose gel Formaldehyde agarose gel Polyacrylamide gel Gel pretreatment Depurination, denaturation, and neutralization - - Blotting method Capillary transfer Capillary transfer Electric transferDot blot has several uses. Title: Northern blotting 1 DNA (or RNA) samples loaded into wells 2 (No Transcript) 3 Ethidium bromide is fluorescent in UV light 4 Northern, Southern, Western In the 1970s Ed Southern of Oxford University invented a revolutionary DNA blotting technique. Reverse allele specific oligonucleotide assays provide a robust method for the molecular characterization of high-mutation spectrum disorders. Southern Blotting: Developed by E. •A northern blot is almost identical to a Southern blot, but it involves the detection of RNA instead of DNA. 2004;4:12. Electrophoresis 23, 2373–2376 Kakita, K. The technique follows the steps: electrophoresis of cell extract (protein) in polyacrylamide gel; blotting of proteins on to nitrocellulose filter paper; hybridization of proteins by using radiolabelled antibodies (I 125 antibodies) of known structure; and detection of hybridized sequences by autoradiography. Southern hybridization commonly known as southern blot is a technique employed for detection of a specific DNA sequence in DNA samples that are complementary to a given RNA or DNA. It is used to. . Biochem. A qualitative and semi quantitative data can be. If the radioactive label is not used, the bound antibody. Whatever system is used, the intensity of the signal should correlate with the abundance of the antigen on the membrane. (1982. 2–2 μg. DNA is restricted with enzymes 3. Dot blot is a simple and rapid technique for the detection of proteins. 23. Use dot plots to present western blot quantification data. The quantity of the specific target is then determined. Here, I describe a simple protocol to quickly check the presence of recombinant protein in cell cultures using a dot-blot experiment. Reverse allele specific oligonucleotide assays provide a robust method for the molecular characterization of high-mutation spectrum disorders. Southern blotting combines transfer of electrophoresis -separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization. The difference lies in the visualization process. BLOTTING • TRANSFER PROCESS: – Capillary Method (Wet transfer. Southwestern Blotting Assay. Dot-blot hybridization method definition, Principle, steps, advantages A simplified 3 minute videoThe video explainsDot-blot hybridization Definition: This t. Follow. 5, 3. DOT BLOT 10. However, one disadvantage of these methods is. An overview of western blot analysis covering protein separation, transfer, blocking, buffer formulations, primary and secondary antibodies, and detection procedures. , O%Connell, K. Southern BlottingSouthern Blotting In 1975 Edward Southern developed this technique that is widely used to detect fragments of DNA . Dr. •This method is also known as DNA blotting/Southern hybridization. The technique is known as DNA transfer or 'Southern blotting' Professor Sir Edwin Southern. Blotting Techniques Kurgat Gilbert 160. Western blot experiments typically include a small number of observations. IMMUNOBLOTTING • Viral antigens are detected with a polyclonal or a MAb onto nitrocellulose paper. Denature DNA 5. Midland Blotting: A Rapid,. ANALYSIS & IMAGING This is the last & major step of the western blotting technique. Southern Blot It is used to detect DNA. 3 Dot Blotting/Slot Blotting: Vacuum Filtration Method 30 1. Western blotting is an important analytical technique used in cell and molecular biology for last four decades. • A Southern blot is a method used in molecular biology for DNA analysis. pptx. ADVERTISEMENTS: Blotting technique is an extremely powerful tool for analyzing gene structure and used to study gene expression, once cloned cDNA is isolated. Open in figure viewer PowerPoint. Nucleic acid labeling and hybridization on membranes have formed the basis for. Northern & Southern Blots. 5. labeling techniques for nucleic acids. Variable volumes of a BSA solution (2 µg/µL) containing indicated BSA amounts were spotted onto a nitrocellulose membrane. Rock on setting 2 for 5 to 10 minutes. The technique was named after its inventor, Edward M. Analysis Expression of ER, (HER-2/neu), and PR with Immunohistochemistry. In the immunoblot, antigens coated on membranes are used as a solid phase in order to detect specific antibodies in patient samples. Whether the DNA sequence is present or not. Western blotting products, and to search our catalog of over 14,000 antibodies . Southern Blot It is used to detect DNA. This is Part B, Blotting Techniques, under the module topic, Nucleic Acid Hybridization & Expression Analysis. 1- The first step in a western blotting is preparing samples: The samples are prepared and loaded onto a gel. The transferring medium then goes through several chemical and physical treatments. views•. 132K views•20 slides. DOT BLOT TECHNIQUE This technique is used to detect the presence of a given sequence of DNA/RNA in the non- fractionated(not subjected to electrophoresis). Fig. The obtained results indicated 100% accuracy when compared with direct DNA-sequencing and gap-PCR (clinical diagnosis kit). This step is known as probing. Dot blotting allows for the rapid screening of a larger number of samples and/or targets than more traditional methods, such as a western blot or in-tissue-based methods. , nitrocellulose) and. Rajanathan. VACUUM DRIVEN BLOTTING Vacuum Blotting (Olszewska & Jones 1988), has several advantages over capillary or. Electrophoretic as well as non-electrophoretic transfer of proteins to membranes was first described in 1979. of MBB&B, CCS Haryana Agricultural University, Hisar Lecture Title: Techniques used in Molecular Probing Agarose or Polyacrylamide gel electrophoresis • When genomic DNA extracted from any tissue of a plant or animal species is digested with a restriction enzyme, it is cleaved into segments. Physiol. The hybridization procedure is the same as in original blotting techniques. Title: Northern blotting 1 DNA (or RNA) samples loaded into wells 2 (No Transcript) 3 Ethidium bromide is fluorescent in UV light 4 Northern, Southern, Western In the 1970s Ed Southern of Oxford University invented a revolutionary DNA blotting technique. J. 2015;1334:85-99. We have applied the reverse dot-blot method to the detection of the beta-thalassemia mutations of African-Americans. 2 ) DNA fragments are blotted onto a strip of nitrocellulose or a nylon membrane. INTRODUCTION 4. Definition: • The enzyme-linked immunosorbent assay (ELISA) is a common laboratory technique which is used to measure the concentration of an analyte (usually antibodies or antigens) in solution. Sample sizes in blot experiments are rarely large enough to justify the use. Dr. SOUTHERN BLOTTING The technique was developed by E. During the dot blotting, the electrophoresis of the protein. The goal DNA is then connected to a molecule with the intention to resource detection. Blotting Transfer of DNA, RNA or Proteins, typically from a electrophoresis gel to a membrane e. Blotting technique including Southern , Northern and Western blotting Rohit Mondal. The size and position of precipitation bands provides the same type of information about antibody amount as the double immunodiffusion method. 2- Transfer (blotting)- Cross-Linking. 93. Dot and slot blot ppt. The reverse dot-blot (RDB) technique is one of the most widely used techniques to diagnose CF[11. Second, dot blot allows researchers to estimate the target protein concentration in a. Rinse the blot with 2× SSC and remove excess solution. 23, and 0. Eastern blot is primarily performed to detect the. in protein blotting experiments, and offers troubleshooting tips and technical advice. RNA dot blot is semi-quantitative technique for rapid quantification of relative concentrations of RNA, developed by Kafatos et al. Blotting technique including Southern , Northern and Western blotting Rohit Mondal 4K views • 36 slides Northern & Southern Blots presentation. Southern blot is a method commonly used in molecular biology. For the quantification of certain proteins of interest within a complex sample, Western blot analysis is the most widely used method. Introduced by Towbin, et al. Southern Blot. Dot-blot hybridization provides a simple, easy, and fast way to quantify the relative concentration of ANF mRNA within cardiac tissue. Separated by electrophoresis 4. DNA or RNA are usually transferred and immobilized to nitrocellulose or, more commonly, to nylon membranes. If the sample is DNA, then it is known as a Southern blotFig 2 highlights several western blot image display practices that can omit information necessary to interpret western blots, like narrowly cropped blots to display only the band of interest [7–11], omitted molecular weight markers [9,10], and missing or poorly used molecular weight labels [8,10,11]. Southern • dna-dna hybridization is the basis. Dot blot is a technique for detecting, analyzing, and identifying proteins, similar to the western blot technique, but differing in that protein samples are not separated electrophoretically but are spotted through circular templates directly onto the membrane or paper substrate. W estern blot is the analyti cal technique used in molecular biology, immunogenetics, and other molecular biol ogy to detect specific proteins in a sample of tiss ue homogenate or extract. Blotting is used for a selection of recombinant plants . Shawn Mad • 315. Complementary single-stranded. f Principle of Western Blotting. Blotting techniques includes southren,northern,western and dot blotting. Bio-Rad's 0. Use dot plots to present western blot quantification data. pptx Maduni3. This technique is the first nucleic acid technique that is developed in 1975 by E. The appropriate amount of RNA for each dot is between 0. Southern blotting is a process that transfer the DNA fragments that separated by electrophoresis into a nitrocellulose membrane/ nylon. tuberculosis lineage identification. The Northern blot, also known as the RNA blot, is one of the blotting techniques used to transfer DNA and RNA onto a carrier for sorting and identification. The subprotocols 4, 5 and 6 describe direct spotting of RNA on to a membrane filter (dot blot). Blot dry on a piece of whatman. BLOTTING TECHNIQUE p-1. It is traditionally used to detect low amounts of proteins in complex samples or to monitor protein expression and purification. This method is also called “immune blotting” because of its nature to use an antibody for specifically identifying its antigen and also protein blotting. , antibodies or nucleic acid probes) to identify the presence or absence of. Blotting is generally done by letting a mixture of DNA, RNA or protein flow through a slab of gel. In Southern blotting, DNA is transferred from a gel to a membrane for hybridization analysis. 3 Reverse Dot Blot. CONCLUSION 11. Blotting Techniques PPT. Nucleic acid labeling and Hybridization on membranes have. Blotting techniques includes southren,northern,western and dot blotting. Abstract. • Blotting allows researchers to find one protein among many, like a needle in a haystack. The dot blot procedure is a well-validated technique, and its use is an alternative to avoiding the protein separation process although the equipment available on the market requires the use of a larger amount of the sample, the amount of sample is between 50 and 100 microliters. B Western Blotting. Bertorini MD, in Neuromuscular Case Studies, 2008 Screening for Known Mutations. • The northern blot is a technique used to study gene expression by detection of RNA in a sample. 18. T. Southwestern blotting is a technique used to study DNA-protein interactions. Northern blot It is. Despite the advantages and disadvantages of different techniques, the routine method employed in the. Southern hybridization Transfer buffer. g. TYPES OF BLOTTING TECHNIQUES BLOTTING TECHNIQUES Southern Blot It is used to detect the DNA. 1. 3. The technique uses three elements to accomplish this task: (1) separation by size, (2) transfer to a solid support, and (3) marking. Blotting Technique Aman Ullah. Block the membrane with 5% dry milk in TTBS (50 mM Tris, 0. Blotting techniques are used to transfer DNA or RNA fragments or proteins from electrophoresis gel to a nitrocellulose sheet or nylon membrane as blotting paper is used to blot ink. The workflow of western blot assay. Instead, the sample is applied directly on a membrane in a single spot, and the blotting procedure is. In this technique, the DNA is cut with suitable. 5K• 31 slides. Bar graphs are the least informative way of presenting continuous data as they conceal the spread of the data and sample size [15,32]. Dot Blot Hybridization Technique: Definition, Principle, Procedure and Applications. 1. txt) or view presentation slides online. Cont…. Kashyap karthikeyan M. doi: 10. . It was first described by Towbin et al. Reproducibility test: binding of preformed antibodies of one human blood donor to three porcine wild-type (WT 1-3), three porcine GGTA1 knockout (KO 1-3) and one human (H. 8% for examination of H&E-stained tissue sections. Despite some limitations such as long time, different applications of WB have not been limited. 1The Oxford College of Science SL NO. PCR products were visualized in 1% agarose gels and purified using the GFX PCR DNA and gel band purification kit (GE Healthcare, Buckinghamshire, United Kingdom). Southern blotting combines transfer of electrophoresis-separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization. Far–western blot analysis is an alternative method to analyze protein–protein interactions that entails probing proteins that have been separated by gel electrophoresis with tagged bait proteins, and then detecting the interacting proteins (prey) through multiple methods. 1K views. Dot blotting is a molecular biology technique used for the qualitative detection of specific proteins, DNA, RNA, or other molecules in a sample by applying small drops or "dots" of the sample onto a solid support membrane and probing it with specific molecules (e. • This requires 1 ) Separation of DNA or DNA fragments by agarose gel electrophoresis . WESTERN BLOTTING 4. 12 μg) were prepared and separated by electrophoresis. Proteins are separated based on their size on polyacrylamide gel. Blotting techniques. Saba Ahmed M. Slot or dot blotting is a technique whereby nucleic acids can be applied to a solid matrix, unfractionated, using a vacuum manifold. 4. 5K•17 slides. Number. Phil. Extract & purify DNA from cells 2. ELISA By: Dr. Nucleic acid hybridization is a very potent technique that can be used for the identification of DNA and RNA species with varying degree of homology and for the estimation of relative amounts of nucleic acid with known homolgy. A dot blot (or slot blot) is a technique in molecular biology used to detect proteins. Keywords: Alpha chain; deletion; molecular techniques; non-deletion; thalassemia. labeling techniques for nucleic acids. Sandwich blot between two pieces of dry whatman and bake in vacuum oven for 1 hour. The DNA detected can be a single gene or it can be apart of a larger piece of DNA such as viral genome. NorthWestern Blot 6. 2. Southern blotting. Dot blot is a technique for detecting, analyzing, and identifying proteins, similar to the western blot technique, but differing in that protein samples are not separated electrophoretically but are spotted through circular templates directly onto the membrane or paper substrate. Similarly, this technique had been successfully applied for the detection of mutations related to resistance to RIF and shown a sensitivity of 91. 88, 0. Comparison of Southern, Northern, and Western analyses of Gene X.