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If youre working with a challenging plant species, such as cacao and cannabis, try out our state-of-the-art BashingBeads for more complete lysis and more consistent yields (Figure 2).11-14. DNA Purification | DNA Extraction Methods | Promega - Promega Corporation QLnoSc3 TuAWbAJ7$ (5hiHFn%wCEJ}=PbcSB#hN(YYXl69Dr ]\79/SiSM J,QOY5#^A@r|)(' g@9_d[v9|L.Th*9I5D(Y J w9K|F>.-c#0G{3F)2jSV7\snE>rl gCe:"6zkO( Purvi Jain - Perfumer - Sawai Fragrances | LinkedIn trailer stream 2.8. Reagents 1. Fast ID is able to extract DNA from a wide variety of sample types. 5- Washing . Other chemicals required in the process are SDS, protease enzyme, Phenol, chloroform, isoamyl alcohol and ethanol. Activate your 30 day free trialto unlock unlimited reading. CTAB extraction buffer, the detergent captures the lipids and the proteins allowing the We have used this procedure successfully with Pseudomonas stutzeri strains which did not give good DNA preps with SDS-based DNA extraction methods due to polysaccharide production. Let us see the protocol. In principle, in the extraction process, CTAB associates with the cell wall polysaccharides and proteins and co-precipitates it along with DNA. 4. Click here to review the details. Sorry, preview is currently unavailable. Activate your 30 day free trialto unlock unlimited reading. DNA extractions using the NGS protocol with (D) no PVP, (E) 1% PVP, and (F) 4% PVP. Now let us come to our major player- CTAB. Instant access to millions of ebooks, audiobooks, magazines, podcasts and more. In this sense, several studies have reported the efficiency of using cocoa leaf material for DNA extraction, however, has not been considered leaf developmental stage as an important variable for obtaining high quantity and quality of DNA. startxref Spin at top speed in microfuge (cold if possible) 10 min, drain off ethanol, retain pellet. DNA must be purified from cellular material in a manner that prevents degradation. [*ZeJ3"P|7He ]nBx)rgrRSfh3sVPkYQxs811Bwiw"gML xMCs34BA^Co{Ba[*- 8`EAr%EVCq)5U J)\)`.7 Use of phenol/chloroform also generates organic waste which requires special storage containers and disposal procedures. ?1tr XE.8TC!qC^t5W3C"w+| 4(XH@CaqB"'1"&)l7%$,Y$DNlNsRF =WPj'Y~u[.qA$D fd9AV'qSFD#>-JBpl W:LlC}P?.EBl!|J)GaABAUoR=$SXjg!K TQQcDxJ$Jh>64=6S[*z~0(OL ORGANIC METHOD OF DNA EXTRACTION 2. The DNA extraction methods used were Cetyltrimethylammonium bromide (CTAB) and Commercial Kit (Geneaid; Genomic DNA Mini Kit Tissue), with variations in the number of nematodes: 1, 5, 10, 15 and . Extraction of DNA from Infected Cassava and Banana from Cameroon . CTAB is used in this method of DNA extraction. MATERIALS: CTAB extraction buffer. overview before delving deeper into the procedure in a step-by-step approach. The solid phase protocol listed below is an alternative. Grind the tissue in a mortar and pestle and add Urea buffer (which contains Urea, Tris, EDTA and NaCl). H|W~qbqh%oa7_$? H A solution of phenol/chloroform/isoamyl alcohol is used to extract plant DNA from cellular debris and once added and vortexed, the mixture separates into three distinct phases: aqueous, interphase, and organic phase. SDS (sodium dodecyl sulfate) is an excellent anionic detergent that can lyse proteins. Then the amount of phenol and chloroform used is greatly reduced (you can use 500 l instead of 5 ml ! Access Free The Cell A Molecular Approach Geoffrey M Cooper - ahecdata.utah CTAB Protocol for Isolating DNA From Plant Tissues, the cetyltrimethylammonium bromide (CTAB) method, various modifications for different plant species, development and optimization of a one size fits all CTAB protocol difficult. Nonpolar molecules migrate into the organic phase and leave behind DNA and other polar molecules in the aqueous phase. conditions, the nucleic acid do not precipitate, and proteins can be removed by extraction of the Activate your 30 day free trialto continue reading. 0000000016 00000 n | DNA Extraction Method 2. Enter the email address you signed up with and we'll email you a reset link. Chloroform is toxic by inhalation and will also damage eyes. separation of the aqueous and organic phases. Boiling Extraction Method VS Commercial Kits for Bacterial DNA Garfinkel AR, Otten M, Crawford S. SNP in Potentially Defunct Tetrahydrocannabinolic Acid Synthase Is a Marker for Cannabigerolic Acid Dominance in Cannabis sativa L. Doyle J, Doyle J. PDF Bacterial genomic DNA isolation using CTAB - DOE Joint Genome Institute Plant DNA Extraction using CTAB Buffer - Genetic Education Ch05 - Chapter 05 solution for Intermediate Accounting by Donald E. Kieso, Jerry J. Mix vigorously and incubate in water bath at 65C for 1 hr. Washing and elution- By alcohol and TE (Tris-EDTA) buffer. Spin for 5-10 min at 13,000 rpm. The DNA pellet do not dry and dissolved immediately in 300 l 1xTE, pH 8.0 at 55C for 5-10 minutes. c*f(M=;"]txGyFUHIEW8[WE%:8ioGSTzPF*G@ ioE EDTA is a chelating component that among These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In this case, it is recommended to add an isopropanol precipitation step after the first chloroform extraction, then do a 70% ethanol rinse, dry pellet, redissolve in 1-2 ml TE. step (Lysis of White blood cells, WBC) 3- Phase . Under these ice-cold ethanol (1000 l) to precipitate DNA incubate at -20C for at least 1 hour. choosing a selection results in a full page refresh. zWf5 rGG]]no|/ Conventional methods of DNA extraction The protocol detailed here is a modification from Saghai maroof et al. CTAB protocol for the isolation of DNA from difficult tissues (high levels of secondary metabolites or polysaccharides), herbarium and soil. Start with sample preparation, clean the tissue and prepare it for homogenization. Evaluation of five genomic DNA extraction methods for downstream UC - Davis Safety Services website: https://safetyservices.ucdavis.edu/safetynet/safe-use-of-phenol. RNA aptamers, specific for different proteins, have been exploited as biorecognition elements to develop specific biosensors (aptasensors). Lysis: Tissue grinder and use detergent. Once the nucleic acid complex has been Dr Tushar Chauhan is a Scientist, Scientific-writer and co-founder of Genetic Education. There are no repetitive and lengthy phase separation steps or hazardous reagents used, so you can further streamline your labs operations and protect the safety of key personnel. Main Outcome Measures: Successful extraction of DNA Results: PCR, RFLP and DNA sequencing gave the expected results in all the DNA samples extracted by all the three methods (boiling, microwave irradiation and the commercial kit). Corpus ID: 102206113; Comparison of Four Methods of DNA Extraction from Needles of Pinus tabulaeformis @article{Ming2012ComparisonOF, title={Comparison of Four Methods of DNA Extraction from Needles of Pinus tabulaeformis}, author={Li Ming}, journal={Hubei Agricultural Sciences}, year={2012} } It actually challenges our knowledge to optimize things in our lab. Redissolve pellet in 100 l EB. Caution must be taken when working with liquid nitrogen for the first grinding step as it can rapidly freeze skin tissue and cause cold burns even with short exposure. After pouring the alcohol off the pellet and drying, the DNA can be re-suspended in a buffer such as Tris or TE. However, in practice, the protocol has several drawbacks: its lengthy, tedious, and low throughput, with many steps that require careful handling, exposure to hazardous chemicals, and several other technical considerations. DNA extraction is a process of isolating DNA. Ideal lysis procedure is rigorous enough to disrupt the complex starting material (plant The Fast ID Genomic DNA Extraction Kit is designed for whole grains, fruits, and vegetables, and the Fast ID Ultra DNA Extraction Kit (Catalog No. DNA was isolated from leaves of 10 plant species ( Cuminum cyminum, Vigna aconitifolia, Pennisetum typhoides, Tecoma stans, Lycium barbarum, Anogeissus acuminata, Tecomella undulata, Zizyphus mauritiana, Phoenix dactylifera, and Eruca sativa) and a fungus ( Fusarium oxysporum) using the CTAB method. Let me know if it works should you try it though! This protocol originally came to us from Evelyne Derelle at Observatoire Ocanologique, Banyuls sur Mer. To browse Academia.edu and the wider internet faster and more securely, please take a few seconds toupgrade your browser. The DNA from leaf samples of agarwood-producing plants was isolated using the CTAB method (Gholibeigian 2021) The matK, ITS, and trnL-trnF gene fragments were sequenced and used to confirm the . After the clear pellet appears, add 500L of TE buffer or elution buffer to the pellet and dissolve the DNA in it. How to Extract DNA From Plant Leaves & Seeds | ZYMO RESEARCH 3. Prepare a working solution from stock. All Rights Reserved. EB wont give as much protection as TE, which also protects against nucleases. We've updated our privacy policy. After a considerable amount of time, the DNA may eventually float to the top of the alcohol layer. In this paper, most frequently used techniques for DNA extraction from insect with different size and condition are outlined. The Graduate's Guide to Synthetic Biology. By whitelisting SlideShare on your ad-blocker, you are supporting our community of content creators. "Processing with GMO detection method and confirmation of results for corn, soy, cotton and tomato."-Microorganisms' cultivation.-DNA extraction (CTAB ptotocol) -Method confirmation according to ISO 17025.-PCR (Polymerase chain reaction) . So the first thing one should have to keep in mind is that depending upon the constituent of a plant cell wall, and the type of polyphenols they contain, prepare a buffer system. Cell preparation and extraction techniques. CTAB* is a strong detergent, it will damage eyes and irritate skin. Handbook of Molecular and Cellular Methods in Biology and Medicine, Third Edition, Techniques in Molecular Systematics and Evolution, Methods in molecular biology (Clifton, N.J.), Microsatellites for ecologists: non-radioactive isolation and amplification protocols for microsatellite markers, Molecular genetic analysis of recruitment and dispersal in the intertidal procelain crab, Petrolisthes cintipes, Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics, Biosensors for RNA AptamersProtein Interaction, Construction of plant bacterial artificial chromosome (BAC) libraries: an illustrated guide, 11 Deep-sea Thermococcales and their Genetic Elements: Plasmids and Viruses, Methods in Molecular Biology TM Methods in Molecular Biology TM PCR Protocols SECOND EDITION Edited by PCR Protocols SECOND EDITION Edited by. Genomic DNA extraction and amplicon generation. This protocol is my own, we used it in our labs during our masters. x$)FcD2k@ c@5#3Q`e^KDzqjpcm|;lm+ou>oyu?mei{:.||vtz~mG18zh]w \v?o.?;]wWv:auaw:t@-Og8ArNg(quz|JFv\'nC~w^P2uwpvRwghEPC"w;x@wb:^Q$~P9&*RlJAWfb:wT]8PJ\*u;Tut7.TJOD$F:ZTi/=l6QMoT 7j!Fq6bIXuO07:Ro2St~Ovt4',b6bJQ\v:"E Add 400 l TE and 100 l NaCl (5 M) to pellet, resuspend cells by vortexing. 1980; 8(19); 43214325. x\KsFW1XD$I~,5>x,}HeVVwmk-m?X/VW|\ Phenol, chloroform and isoamyl alcohol DNA extraction. Other methods of DNA extraction include salting out , cesium chloride density gradients , and chelex 100 resin [16, 17]. Invert tube on paper towel with lid open for ~5 min to drain last bits of 70% ethanol, then transfer to 60C incubator (OK if tube is lying on its side, but lid open) for 10 min to dry residual ethanol. %PDF-1.4 % Liquid nitrogen prepares the fine power of tissue and also deactivates nucleases by providing an extra chill environment. Various chemical constituents of DNA extraction buffer allow effective isolation. xref abyssi strain GE5. For instance, the DNA extraction setup and buffer which works for animal tissue, cant work for plants. By centrifugation, such co-precipitates can be removed in consecutive steps.The structure of CTAB. Weve updated our privacy policy so that we are compliant with changing global privacy regulations and to provide you with insight into the limited ways in which we use your data. It appears that you have an ad-blocker running. Centrifuge the sample at 25,000rpm for 5 to 8 minutes and transfer the supernatant into another tube. EB (Elution buffer): 5 mM Tris-HCl. 1. Decant and dry the pellet at RT. determine the quality and yield. Aboul-Maaty NAF, Oraby HAS. Increasing the heat and duration of the CTAB step might help with tough cells (eg. ^ cQmG;Q5h7~S.KC10NdXwkx|%!UPvDPXN2_82q?Lz>O&=pq7^9vmA(PdLe m C1!j 3SK97vU|(TgEw'q>jh:k"a6==UDnMW+6i4~t5F&CfYn*OGB"GrL#~]mT$zUIa 6/iDbe h'*omlb_i*G'*MBP q08C q~x]ufAi=Y a3FanN(CSQY6YvQ*K=($b6(+R@6}IDBDD+MD]R@+?%|C,Hi C}dFf13WI ve^*G. Tissue grinding can vary between samples, leading to significant variation in extraction efficiencies and quality of DNA. Add 500 l phenol:chloroform, vortex until milky solution obtained (~20 sec), spin at top speed in microfuge for 5 min, retain supernatant. These recognition elements have been coupled to piezoelectric quartz crystals and surface plasmon resonance (SPR) devices as transducers. This is because the base pairs in DNA absorb UV light, therefore amount of bp is directly proportional to DNA concentration. Now customize the name of a clipboard to store your clips. 1- Lysis . High molecular weight DNA yield in the range of 328 to 4776 ng/L with an average . Modified CTAB Technique for Isolation of DNA from some Medicinal Plants Safe Use of Phenol | Safety Services. These two solvents are generally considered hazardous. I hope to test this at some point to confirm some of these comments, but as my source for many of these protocols, I know he is rarely, if ever wrong. In contrast to the original method, the modified CTAB procedure is faster, omits the selective precipitation and CsCl gradient steps, uses less We've updated our privacy policy. PK ! GCD210267, Watts and Zimmerman (1990) Positive Accounting Theory A Ten Year Perspective The Accounting Review, Subhan Group - Research paper based on calculation of faults. purpose, the aqueous solution is first treated absolute ethanol or isopropanol. DNAITPCRDNA Our binding chemistry, wash solutions, and spin column technology remove polysaccharides, lipids, and other common downstream inhibitors and contaminants providing ultra-pure DNA, with minimal loss. To overcome the challenges presented by plant tissues, the cetyltrimethylammonium bromide (CTAB) method has become the go-to protocol for DNA extraction and purification from leaves and seeds. First report of 'Candidatus Phytoplasma australasia' related strain XeQ You can download the paper by clicking the button above. To make things even more effective use RNase as well in combination with proteinase K. Proteinase K and RNase are commercially available. Romero Navarro JA, Phillips-Mora W, Arciniegas-Leal A, et al. Cell wall and nuclear membrane lysis- by chemical treatment, tissue homogenization using liquid nitrogen and physical grinding. *e *S{(h-IB{Zec>]K pwbhYP=[! By binding Mg with Instant access to millions of ebooks, audiobooks, magazines, podcasts and more. PRINCIPLE: The extraction of genomic DNA from plant material requires cell lysis, I will also explain variations and concentrations of different CTAB buffers that you can try and use. endstream endobj 38 0 obj <> endobj 39 0 obj <>/Font<>/ProcSet[/PDF/Text]>>/Rotate 0/Type/Page>> endobj 40 0 obj <> endobj 41 0 obj <> endobj 42 0 obj <> endobj 43 0 obj <> endobj 44 0 obj <> endobj 45 0 obj <> endobj 46 0 obj <> endobj 47 0 obj <>stream PPT PowerPoint Presentation I .g$!=n]BQ|nTr|0T5t4@XIR$fwyVIszC9Er(ze*RzoI$M%8FM=Mi fR{X[:[F R=XFBdvk>*H7&NIP'B.4GsXUS6j'^p2+#hY[xBYUJuk (L)4/vB`])tS2\YI!MSCKVN",NAN}/DKHFR apart, the purification of DNA is performed. By accepting, you agree to the updated privacy policy. 10 Different Types of DNA Extraction Methods (Updated), 7 Ways to Determine Genotypes Using Gel Electrophoresis, DNA Extraction From Dried Blood Spot Samples: Protocol + Comprehensive Guide, Metagenomics Made Easy: Streamlining DNA Extraction With Bead Beating, A Guide To Next-Generation Shotgun Sequencing In Metagenomics: Technique, Advantages and Challenges. )Z$m|h_8"dOtv#P%w;:K&z]x.A`5.AZ%q9;eU! The basic principle of DNA isolation is disruption of the cell wall, cell membrane, and nuclear membrane to release the highly intact DNA into solution followed by precipitation of DNA and removal of the contaminating biomolecules such as the proteins, polysaccharides, lipids, phenols, and other secondary metabolites by enzymatic or chemical A protocol for high-quality genomic DNA extraction from legumes Click here to review the details. oBrgc;ntf24t``D7Sw8C&NQn"1_* ?dNOHhc/s3 ^D{6n>`SFcMedqW%MT0P!X?K` FlThc0 'wU/n;@wP~0ypKO8gW>LKn:?~hlfTvU=a"or^FuP]I.u;H\"X3Mh]buUbWQ5,N,0P+A+SXEAZV:57{F:999(G!Eh#`Ln{abDH"{JKiBH;jA} }9\cY"1La Isolating plant genomic DNA without liquid nitrogen | SpringerLink Nucleic acids research. We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract nucleic acids from plant tissues. Care must be taken while handling CTAB as it has corrosive and toxic properties. CTAB DNA extraction is a simple, effective, cheap and reliable method to extract DNA from plants, especially for RAPD analysis and barcoding. 10 Different Types of DNA Extraction Methods (Updated) - Genetic Education Find out more Dismiss The use of Urea is the easy and safest way to extract DNA from plants and sufficient enough for routine PCR and restriction digestion but not enough for sequencing or microarray analysis. We use cookies to track visits to our website, and we don't store any of your personal details. DNA EXTRACTION: 5. ^ 0=pLRto6Q&N(Na*Y;"I$1k*wkX 5|AQ>4DWii/:&"E N CTAB solution: 3% CTAB, 1.5 M NaCl, 10 mM Na 3 EDTA, 0.1 M MOPS-acid; If that all seems like a bit much, you arent wrong. Extraction of DNA, RNA, and protein is the basic method used in molecular biology. The cells in a sample are separated from each other, often by a physical means such as grinding or vortexing, and put into a solution containing salt. 0000001976 00000 n A lysozyme incubation can be added if cells dont lyse well with CTAB alone. However, other chemicals like RNase, proteinase K, SDS and PCI may be used to improve the quality and yield of DNA. membranes (such as those around the mitochondria and chloroplasts) have been broken 4. Magnetic bead-based DNA extraction: 8. So these three things (cell wall composition, secondary metabolites and turgor pressure) need advanced treatments. CTAB Protocol for Isolating DNA From Plant Tissues This protocol can be used on most gram-negative bacteria to obtain high-quality genomic DNA, which is both pure and very high molecular weight. Add liquid nitrogen to the tissue and roughly grind the sample into the mortar and pestle. Isolation of good quality DNA is a fundamental first step in insect DNA barcoding which is obtained by standardizing the DNA isolation method. Institute of Biomedical and Genetic Engineering (IBGE), Dna fingerprinting matreilas & methods of chilli, Dna extraction from fresh or frozen tissues, Human biosafety issues of nanomaterials used as nanomedicine, Introduction, pathophysiology and treatment options for covid 19, Inhibition of Gene Expression at RNA Level. It was adapted from Winnepenninckx B. et al., 1993, TIG: 9 (12), 407 (Technical Tips). % DNA was extracted directly from freshly harvested leaves ground in pre-heated CTAB buffer. Pre-prepare a CTAB solution (10%) by heating and store it at 37C temperature. Plan your day carefully and set aside the proper amount of time to complete the entire protocol. 3) CTAB is widely used to disturb membranes and common method used in Plant DNA Extraction whereas SDS is used to solubilize membrane and used in Bacterial DNA Extraction, remove. Heating at 50-60C (up to an hour, intermittent mixing) and/or addition of more EB may be required to dissolve all the DNA. Government Pharmacy College Sajong, Government of Sikkim, University of Horticultural Sciences, Bagalkot, Endosperm culture and somatic embryogenesis, Biotechnology experiments 2nd semester (LNMU Darbhanga), Genomic Dna Isolation From Blood, Bacteria and Plasmid DNA Isolation, Lectut btn-202-ppt-l16. Rl%= PPTX PowerPoint Presentation We use this method for extracting genome sequencing quality (i.e. Euphrasia nankotaizanensis (Orobanchaceae) is a rare alpine herb that is endemic to Taiwan. A qPCR Assay for the Fast Detection and Quantification of Colletotrichum lupini. Alex: Gee, thats a lot of chloroform, might be a while before I risk trying this protocol. << /Length 1 0 R /Filter /FlateDecode >> DNA extraction is a process of isolating DNA from any biological cell to use it for analysis like PCR, sequencing or restriction digestion. Angeles JGC, Laurena AC, Tecson-Mendoza EM. It will surely make things easy. Under these 10% w/v CTAB (Cetyl trimethylammonium bromide), Autoclaved. This method will help the researchers to efficiently isolate DNA from . homogenate to sterile Eppendorf tube. Load 2-4 L of isolated plant genomic DNA in 0% agarose gel and You choose the most appropriate method ! Ethanol and NaCl are used to remove plants polysaccharides. This protocol is wholly adapted from the Coleman Lab protocols, including notes and comments. 6R(D$2(>1bD44b9!nGH-NCgVw IIOCebfcDKpC]rY8& "7 k@- Main steps in DNA isolation procedure: 4 Lysis Removal of proteins and contaminants Recovery of DNA A number of commercial DNA purification kits use the same principles as the manual method. There are five basic steps of DNA extraction that are consistent across all the possible DNA purification chemistries: 1) disruption of the cellular structure to create a lysate, 2) separation of the soluble DNA from cell debris and other insoluble material, 3) binding the DNA of interest to a purification matrix, 4) washing proteins and other Tris-HCl gives the solution a pH Proteinase K is yet another excellent ingredient that also can digest proteins- DNase and RNase present in the cell and protects the nucleic acid from degradation. %PDF-1.3 Also, prepare a 10% stock solution of PVP. Phenol is volatile and can burn the skin. out, while the nucleic acid precipitates. Ch13 - Chapter 13 solution for Intermediate Accounting by Donald E. Kieso, Jerry J. additional purification, or wash, of the nucleic acid from the remaining salt. In this article, we will discuss plant DNA extraction and CTAB buffer. Remember to boil RNAse before use to kill any DNAse in it (see Sambrook for method). CTAB Method DNA extraction - Molecular Biology BCH- Isolation of plant genomic DNA by modified CTAB - Studocu How to extract plant DNA using CTAB method molecular biology isolation of plant genomic dna modified ctab method principle: the extraction of genomic dna from Skip to document Ask an Expert Sign inRegister Sign inRegister Home Application of Genome Wide Association and Genomic Prediction for Improvement of Cacao Productivity and Resistance to Black and Frosty Pod Diseases. DNA extraction is one of the first steps to many research, forensic or clinical analyses. Download our ebook: From DNA Extraction to PCR, Plant DNA extraction needs a CTAB DNA extraction buffer that contains ingredients that can effectively remove polysaccharides and polyphenols. Extraction(CTAB) Buffer 1.4 M Na Cl Breaking cells open to release the DNA. How to Prepare Lysis Buffer for Different Types of DNA Extraction Methods? Slowly pipette out and retain supernatant avoid interface material and lower phase. Q TP? For example, some plants need only simple grinding in mortar and pestle, while some need liquid nitrogen. Among the Thermococcales, the most important order of the phylum Euryarchaeota, only one virus is isolated and completely characterized to date. Plant cell walls are very difficult to break down and the cells contain many compounds that impede extraction and inhibit downstream molecular biology applications. There are also more than 20 steps in the protocol and as the number of samples increases, the amount of time needed to complete DNA extraction increases substantially. Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. 0 (100 mM Tris-HCl pH 8, 2% (w/v) CTAB, 20 mM EDTA, 1 M NaCl, and 1% (v/v) Wash with around 700 L of 70% ethanol. This research evaluated five genome extraction methods including Protocol A (main CTAB), Protocol B (CTAB without -mercaptoethanol), Protocol C (CTAB without ammonium acetate), Protocol D (modified method of Murray and Thompson), and Protocol E (Gene All-Kit), to determine their ability to extract DNA from fresh and herbarium leaves of A. fridae. A total of 30 samples of five algal genus were used in this study. Spin-column DNA extraction: 7. Muhammad I, Zhang T, Wang Y, et al. CTAB Protocol for the Isolation of DNA from Plant Tissues 10 Secret Tips for DNA Extraction to Get Good Results. In addition, the efficiency of PVP increases greatly by adding beta-mercaptoethanol.