Science method
Plant DNA Extraction - Science method
Explore the latest questions and answers in Plant DNA Extraction, and find Plant DNA Extraction experts.
Questions related to Plant DNA Extraction
Greetings to all!!
DNA is isolated from infected cotton leaf.
The image is attached. It looks kind of shearing.
What are possibilities to use it for PCR?

The QIAmp Fast DNA Stool Mini Kit has two main protocols listed in the handbook, accompanied by some simple notes:
1) Isolation of DNA from Stool for Human DNA Analysis
Lysis conditions in this protocol are optimized to increase the ratio of human DNA to nonhuman DNA.
2) Isolation of DNA from Stool for Pathogen Detection
Lysis conditions in this protocol are optimized to increase the ratio of nonhuman DNA to human DNA.
I'm interested in extracting and amplifying plant DNA from mammalian faecal samples to explore dietary habits. From my reading, it seems that both of these protocols have been utilized interchangeably in various papers to investigate diet, however, I'd appreciate any thoughts on whether one of these protocols might be better than the other for increasing plant DNA yield?
Thanks!
Is it feasible to swap 2-mercaptoethanol with any alternative chemical during plant DNA extraction from sediments?
One of my colleague is trying to isolate DNA in order to check a bacterial disease in olive. However she couldnt manage to isolate DNA up to now. She tried plant DNA isolation kit and CTAB buffer but after isolation she couldn't see any band in agarose gel. Also spectro results are not good. very limited amount of DNA and very bad purity. Do you have any experience DNA isolation from olive leaves or vein of the olive leaves?
Hi, I'm trying to isolate total DNA from dried oak leafs using CTAB protocol.
Normally, after the incubation in the lysis buffer (2% CTAB, 100mM Tris, 20mM EDTA, 2.8M NaCl, 1% PVP, 1% BME) and two washes with CI 24:1, I put 500µl of the aqueous fase in a clean microcentrifuge tube, add 500µl of chilled isopropanol + 50 µl of Sodium Acetate and a DNA "cloud" form immediately. Lately the isopropanol mixes poorly with the aqueous fase (some times not mixing at all) and the precipitate doesn't form. Does anyone know what can be happening?
I am looking for a good plant DNA Extraction Workstation and a kit for a 96 well plate. I've already heard about the Beckman Biomek Liquid Handling Station, the BioSprint 96 robot, and also the KingFisher™ System. Do you have any experiences with them? Can I use the same workstation to do PCR prep? About the Kits, do you have any experience with Sbeadex® plant kit, Kleargene Plant 96 DNA kit, MagMAX™, or BioSprint 96 DNA Plant Kits? How is the average DNA yield and quality? Do you have any other equipment and kit suggestions for wheat leaf or seed DNA extraction?
I have a question about the role of NaCl in the DNA extraction process. So for NaCl concentrations under 0.5M, CTAB and DNA molecules can create complexes. In those concentrations, proteins and other hydrocarbons are still soluble in water except the DNA-CTAB complex. If we raise the concentration of NaCl then the complex of CTAB-DNA will be soluble to water too. Correct me if I'm wrong until now.
So I don't understand something. Why do we use NaCl?
Let's assume that we don't add NaCl. Will the CTAB create complex with DNA or not? As I read CTAB creates ionic bonds with DNA (phosphate groups). So in this situation (without NaCl) CTAB must be able to create complex with DNA too.
So why did we use NaCl? Just to keep proteins and other hydrocarbons soluble in the water or there is something else that NaCl helps in this complex creation?
Good day.
Does anyone have any protocol in using disruptor genie for plant tissue lysis? What bead size and amount did you use? How many minutes and rpm?
Thank you very much.
Looking expert opinion...
I have collected marine sponge samples and were shadow dried for two weeks. Now the sponge samples are well dried and can be directly powdered by grinding. I would like to study the sponge associated actinobacterial populations (uncultured) from the dried sample rather than fresh sample.
Here comes my doubt,
If we grind and use the sponge powder for metagenomic DNA extraction, does the DNA be damaged/sheared ?
or
can directly use the dried sponge material (without grinding) for metagenomic DNA extraction?
Kindly, some one clarify my doubts.
Thanks in Advance,
Siva
Hi there,
I'm looking for a High Molecular Plant DNA Extraction Kit for subsequent Nanopore sequencing. Does anyone have experience using commercially available kits? Any suggestion/recommendation?
Thank you very much beforehand :)
I've found that my concentration of DNA is less than the expected value.
Hey everyone. I collected some leaf tissue samples from the plant Phragmites australis from which I am hoping to extra DNA for sequencing. I will be extracting DNA using Qiagen DNeasy Plant Mini Kits. I was in a rush when storing them, however, and I just placed them in ziploc bags in the freezer at -20oC. They had been kept in the same bags in a cooler while transporting them from the field to the lab.
Is this going to be okay? They've been in there for a few weeks at this point and it may be another few weeks or even a month or two before we will be able to begin lab work. Would it be possible to move them to -70oC now or is it too late? Can they be thawed and dried at room temp in silica gel? Just wondering what my options are here and what I need to take into consideration. At this point, it is too late in the season to collect new tissue samples so this is all that I have to work with.
Thanks in advance!
I am preparing CTAB buffer and I read that addition of PVP (Polyvinylpyrrolidone) helps to remove phenolic compounds. Most of the times in protocols there is no additional information about average molecular weight of used PVP, but I found in some, where it was specified that it was PVP 40 (mol wt 40 000) for DNA extraction. But I wanted to know if I can use the same for RNA extraction or should I use PVP 360 (mol wt 360 000)?
Dear fellow scientist,
For RNA isolation I use Trizol, however is pH of Trizol between 4- 6 which allow
DNA to be retained in the organic phase and interface, leaving the RNA in the
aqueous phase?
Greets,
Tse
Please provide me practically viable solution.
I am doing DNA extration from whole saliva using Vivantis DNA extraction kit for Viral nucleic acid purification. After DNA extratcion on PICO drop the values of DNA is 333 microgram per microliter or 423 microgram per microliter, whereas the ratio of A260/A280 is 3.3 or above 3 and ratio of A260/A230 is above 3. Is these values are normal? or high ratios mean?
Is this high ratio of A260/A280 and A260/230 is okay for future PCR analysis?
I want to isolate the DNA from the chloroplast of Brassica napus. In downstream, I will use that DNA to amplify my target region for cloning purposes. Presently I have isolated whole genomic DNA from it. It was thought that this DNA will contain the chloroplast and mitochondrial DNA in it as well and will be amplified by the chloroplast specific primers. But I am not getting any amplification. Now I want to isolate the chloroplast DNA and give it a try with the PCR. Please suggest for me. Thanks
I am looking for reports about Roots expanders used in ORC-systems.
Especially when they are used for steam quality <1.0.
Reports about Roots expanders used in water/steam systems are of course also of interest.
I have extracted DNA from cacao leaf tissue (freeze dried).
I used CTAB buffer (5%) along with 1% B-mercaptoethanol and 3% PVP. I did the DNA extraction overseas and transported it as a pellet to Australia. When they arrived in Australia I stored them at 4 C after resuspending them in TE buffer. However, when I checked the quality on agarose gel surprisingly all of my samples have degraded DNA.
Could anyone pls advice me what can I do to prevent the degradation of the DNA? The downstream application of the DNA will be DarT analysis.
I have attached the gel picture as well.
Regards
Gurpreet

If I have a plant leaf with Alternaria sp. infection. Is there a commercial kit to extract the fungal DNA only? OR the plant DNA ?
I am isolated DNA from some folliose lichens, then purified in ethanol at 13000 rpm, after 15 minute drying, pellet doesn't dissolved in elution buffer. I am also using warm water and ( 65 C) elution buffer but pellet doesn't dissolved after 15-20 days. How can i troubleshoot this problem.
I have to extract DNA from a large number of garlic leaf samples for SSR analysis. I am doing a bead milling technique using SPEX SamplePrep GenoGrinder. My current protocol involves using lyophilized garlic leaf tissue (around 50 mg), two 3.2 mm metal beads, 2 ml tubes, 1200 rpm for 45 sec with pause every 5 sec to rest tubes in ice. My extraction buffer is composed of 3% CTAB, 100 mM Tris-HCl ph 8.0, 20 mM EDTA, 4% PVP, 3% B-me, and 2 M NaCl. My problem is that all of the DNA that I get from my method is sheared/degraded. Is there anyone who has an experience using the bead milling technique for DNA extraction of fragile leaf tissue?
Thank you in advance.
We are working on the functional recovery aspects in response to various phytochemicals. Our targets will be the expression levels of AChR. We want to extract the RNA in muscles but we don't have tissuelyser in our lab. Would anybody advise us the better way for RNA extraction without using tissuelyser please?
It says it is to prevent shearing but does not explain why. Anyone?
As I am trying to extract the DNA from herbarium Fungi and some of them are really old, need some advice to get the best result?
Thanks in advance.
I'm planning to do extractions for metagenomics & metatranscriptomics of insect gut contents. Any other recommendation for quick isolation and separation of DNA and RNA from the same sample would help me. Thanks
Can anyone kindly tell me what amount of rice seedling leaves and root is required for isolation of good quantity of RNA ? Should I weigh the freshly chopped samples before homogenizing with lN2 or after?
Also can anyone please suggest methods for storage of rice seedling root and shoot samples for later RNA isolation?
I want to make sure that my plant extract has no fungal contamination
The same way as universal primers targeting the 16S rRNA gene are used to detect bacterial DNA, which is the best way to assess the occurrence of plant DNA in soil samples? No need to evaluate diversity or taxonomy, just the occurrence.
Hi,
I would like to extract RNA from biofluids, and we have a home-made protocol that I got from a collaborator, that uses Zymo spin columns. However, we have bunches of miRNeasy micro minelute spin columns (Qiagen). Do you know whether those columns are similar in terms of composition, chemistry etc.?
i am working on Gene expression but cant get it done because i am not getting any RNA extracted from my plants. my colleagues says its just because of rapid degradation. what are the main reasons for RNA degradation ?
i am using rice crop..
DNA will be used for SSR detection in coffee.
Thanks.
I am doing some RNA extraction from tomato leave samples for my gene expression work and by checking the concentration of my RNA extracted on a Nanodrop spectrophotometer, the concentration I got were as high as 1000ng/uL.
Can someone please help?
My student has bundles of bee-collected pollen from a phenology study, and would like to get DNA barcode data from them. We have tried following protocols involving liquid N2, and grinding 0.1g of pollen into "fine powder" with mortar and pestle. It never seems to "powderize" but rather turns into a mushy mess. Yields are very low, between 8 and 12ng/uL. Would appreciate any tips on pollen extraction. Post-(semi-successful) powderization, we use the Qiagen DNEasy plant extraction kit.
Hi everyone,
how do you determine the size of genomic DNA? You cannot use a gel because the DNA is too big, any suggestions?
What would you do to determine the size of big DNA extracted from cells without breaking it?
Thanks
I want to ask that I have isolated DNA from bacterial samples but the problem is that i'm not getting any amplification using the same isolated DNA for PCR I want to give it a proteinase k Treatment again but I don't know the exact amount of proteinase k to be used for 25-30 microlitre of DNA
I try already 2 protocols but there are not good enough, the nanodropo and the electrophoresis gel does not give us good results!
How can extract DNA genomic with high purity in fungal with the CTAB method?
In order to get a higher concentration of DNA with Molysis extraction kits (Molzym-Coimplete5 and UMD) we are eluting columns with 50 ul, instead of 100 ul as recommended. Can it affect DNA quality?
Genomic DNA extraction using CTAB I use 800 CTAB buffer and 30 microlitres RNAase.I use Maize plant leaf fresh

Dear all,
As a newcomer, this is my first time to ask for help about my research projects. please say all you know about my question. Thanks very much.
Now I am busy with the isolation genomic DNA from the chocolate, but the results is very poor, the DNA elution is lightly yellow and no target band can been seen by the agarose gel. How to deal with the though question? Do the every member of the researchgate have a ideal method to resolve the problem?
best wishes
Yunjing Zhang
I am doing an rna extraction from tomato leafs, using the Trizol protocol. But when I add chloroform, and centrifuged the samples, the hidrofilic portion gets a pink coloring. And the color keeps going in the steps. Any one know what it is? I am afraid that could be Guanidinium thiocyanate, and could be resulting in a low proportion of 260/230. Any information is valid.
Please I need the most efficient method of isolation of natural dyes from plants.
Any ideas?
I was working with some lignans but accidentally I contaminated these ones with a mixture of alkaloids (the ones I said before). So I was thinking on using recrystallization but I wanna know if anyone of you guys has a better idea or maybe some tips to do the recrystallization.
Thank you!
I am just into mycorrhiza research. I understand that preserving root samples in ethanol is easy and safer, especially for large samples. I would like to know whether the ethanol (70%) must be rinsed with water before the clearing and staining steps.
Other colleagues are extracting good quality DNA of other crops with the same chemical. so all chemicals are working good.
I want to know which of these options apply during fieldwork.
Can the beads be washed with phenol or chloroform, or any other solvent?
I need to remove lactophenol cotton blue from samples for PCR - there is an ethanol wash step in DNA extraction protocol, but perhaps another solvent (phenol/chloroform?) should be used?
100 ul barrier tips is common and needed. However 10 ul barrier tips may not be practically needed. Please give your opinion.
DNA extraction from a plant leaves rich in phenolic compounds.
I have tried following several protocols to extract DNA for my future SSR using primers for PCR amplification (Lodhi et al. 1994; Doyle & Doyle1987), from Sesame leaves. Interestingly, some samples showing good and clear band (down) however, other samples (upper) are not showing sharp band on my Agarose gel electrophoresis. Does anybody know what is the problem with my samples or which protocol to follow for good quality DNA isolation? I would very appreciate for any comments and guidance

Any one can suggest a protocol for silica based plant DNA extraction.
Hi,
I was wondering, if it is possible to extract total RNA and DNA from insect which have been fixed or stored in 70% ethanol at RT and some specimen at 4 degree. The isolated total RNA would be used in RNA sequencing analysis and Whole genome analysis.
If anyone knows information about the isolated total RNA and DNA, is it working for sequencing and genome analysis or not and what type of ethanolic effects showing on specimen RNA/DNA. I would really appreciate for the help.
Thanks in advance,
Maheshkumar
I want to extract DNA from pyralidae moths.
Has any one extracted DNA from the walnut leaf samples preserved in 70% ethanol? I am worried about the quality of DNA. Can it be used for further analysis like linkage mapping and studying of genetic diversity ?
I isolate genomic DNA from dried soy bean by genomic dna isolation kit. But is it possible to occur that DNA concentration is seen when tested by nano drop but band is not seen when it is run by gel electrophoresis? what does it mean?
Extracted DNA from soil by using Nucleospin kit..... followed the procedure and even reduced the samples quantity..but still not pure DNA.
I am trying to find any DNA extraction kit that suitable for field usage. It will be better not to use a centrifuge. Does anyone have any suggestion?
What is best method for total DNA extraction of endophytes from plant tissue? Can i prefer CTAB method for total DNA extraction or any DNA isolation kit? If kit is best then please suggest me kit name.
I am trying to isolate genomic DNA from blood with DNAzol. I am getting degraded DNA. Solubilized fraction is red in colour.
Does anyone know an alternative method to the disruption of seed that does not use liquid nitrogen?
They are very small seeds, beads and blender are not efficient.
Thanks
DNA extraction using CTAB method or a kit.
The sample is either the mesocarp or peel.
I am storing leaves in CTAB buffer before I extract the DA from the leaves. How long can I store (and in what conditions) CTAB buffer before I can use it? I know that when the solution begins to crystallise, it should no longer be used.
Is there anyone have quick DNA extraction protocol?
I use maize as my plant material, and extract DNA by Epicentre' quick extraction kit. But this item is stopped selling now. I am trying to preparing the quick extraction reagent myself. If you know how to prepare the quick extract buffer, please share it with me. Thanks a lot!!
As i tried to extract DNA from fresh plant tissues using kits, or CTAB method DNA was in bad quality.
I had already acetolysed the samples for 2 times, but still no success. I'm dealing with dried pollen and flower samples collected from few herbarium.
What is the actual difference between extraction procedures of plasmid and Genomic DNA
I would like to know the major role of ethanol while extracting dna or rna. In all the solvents used, ethanol has to be added first. Why? Please, I need your help
Greeting to all researchers!
Can anyone suggest me with a successful procedure for extraction of genomic DNA from herbarium material of Red algae (Rhodophyceae).
So far we are successful in extracting genomic DNA from fresh material by SDS method. Since this group consist of high amount of polysaccharides, we find it difficult to extract DNA.
Thank you all
Swetha
Hello,
I just want to find out if you have attempted to recover protein concentrate from the moringa leaf using a green extraction system.
Thanks
I have Whatman UNIFILTER microplates used for DNA extraction from Arabidopsis thaliana and I would like to know if there is a way to clean and re-use them (they are very expensive).
I already using Powersoil DNA extraction kit but the result from the extraction was not good
I've been sing CTAB to try to extract genomic DNA of fungi for whole genome sequencing. I've attached a photo of how most of my samples look. Can you tell me what's going on? I know I have a lot of RNA. I am using purelink RNAse A but I guess it's not working. My question is:
Do I have genomic DNA? Is it all degraded? The first band of the ladder is 23,130 bp. I'm using DNA-HindIII
