Open AccessArticle

Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells

Elizabeth Cristina Vieira de Freitas

¹,

Francisca Alves dos Santos

¹,

Maria Raíssa Vieira Lopes

¹,

Dárcio Luiz de Sousa Júnior

²,

Tássia Thaís Al Yafawi

¹,

Ana Carolina Ferreira Araújo

²,

Priscilla Ramos Freitas

²,

Irwin Rose Alencar de Menezes

^2,*

Henrique Douglas Melo Coutinho

^2,*

and

Maria Karollyna do Nascimento Silva Leandro

Health Campus, Doutor Leão Sampaio University Center, Juazeiro do Norte 63040-405, Brazil

Department of Biological Chemistry, Cariri Regional University, Crato 63105-010, Brazil

Authors to whom correspondence should be addressed.

DNA 2024, 4(4), 417-426; https://doi.org/10.3390/dna4040029

Submission received: 11 September 2024 / Revised: 22 October 2024 / Accepted: 8 November 2024 / Published: 12 November 2024

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Abstract

Objective: The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 × 10⁶ CFU of Candida albicans, targeting to overcome the challenges in the extraction of these genetic materials. Materials and methods: From this, treated silicon carbide (SiC) granules were added to fungal samples from methods 1, 2, and 3 obtained from aliquots of BHI or Sabouraud medium to cause cell lysis and enable the isolation of these macromolecules by phenol and chloroform. The concentration and integrity of the extracted nucleic acids were analyzed, respectively, by spectrophotometry using the A₂₆₀/A₂₈₀ ratios and 1% agarose gel electrophoresis. Results: Therefore, method 3 is the one that most comprises samples considered pure of both DNA and RNA, simultaneously. Furthermore, the presence of intact RNAs corresponding to the base pair size such as 5.8 S rRNA and tRNA was verified during electrophoresis, considering the particularities of RNA, which makes it very unstable and easily degraded. Conclusions: Thus, it results in a faster and simpler method in addition to obtain promising results using minimal amounts of biological sample and offering a valuable alternative for small laboratories to work with molecular biology.

Keywords:

chloroform/phenol; extraction; nucleic acids

1. Introduction

Candida albicans is a commensal fungus that integrates the microbiota of various mucous membranes in healthy individuals without causing disease. However, for immunocompromised hosts, this species can become pathogenic. Accordingly, it has the ability to coexist as a commensal and as a fungal pathogen in humans. The development of various polymorphisms, nutritional adaptation, and biofilm production, among other factors, were used for adaptation and virulence [1,2,3,4].

Therefore, the use of molecular biology based on the study of genetic material is what makes it possible not only to understand particular and fundamental characteristics, but also to detect these microorganisms when they are present in other living beings. Thus, the isolation of genetic material is the initial milestone of any more detailed study of any being. Regardless of their complexity, they have the ability to synthesize macromolecules, such as nucleic acids and proteins, which contain all the necessary information for cellular functioning [5,6,7].

For this reason, there are no single or fixed protocols for the extraction of genetic material since its isolation, quality, and quantity requires a careful choice of the extraction method which can vary according to the samples used. Furthermore, adaptations to these methodologies aim to create less expensive and faster protocols that maintain the quality and yield of the isolated material [8,9,10], making them valuable both for research purposes and for use in laboratory routines that aims at the molecular diagnosis of microorganisms.

Accordingly, methods that use silicon carbide (SiC) in their composition seek not only to assist in the rupture of cellular structures, but also to provide homogeneity to the samples. Thus, SiC granules are stable and inert ceramic compounds produced industrially that can be used as abrasive agents in the cell lysis stage. Since the structures that compose cell walls require more vigorous conditions for the release of cytoplasmic content, they are more resistant compared to structures that have only a plasma membrane [11,12,13,14].

The adaptation is significant as it seeks to optimize the process of extracting fungal genetic material, which is interfered by several factors, in addition to being the first step of molecular biology studies, especially in small laboratories. Therefore, the objective of the research was to evaluate the yield, purity, and integrity of the nucleic acids isolated by the adapted protocol for the joint extraction of DNA and RNA from Candida albicans using a chemical method associated with SiC from some cells, followed by gradual additions to evaluate its effectiveness.

2. Materials and Methods

2.1. Preparation of Materials

To perform the joint extraction of nucleic acids, the method of Oliveira et al. [15] was used, in which all the solutions, including reagents, were prepared in deionized water treated with diethylpyrocarbonate (DEPC) 1:1000 (v/v), at 37 °C for 24 h, and autoclaved. The glassware and accessories used were also treated and the resistant ones were autoclaved at 120 °C for 20 min and the plastics materials were guaranteed to be DNases and RNases free.

2.2. Obtaining and Preparing Fungal Samples

To obtain the fungal samples of the Candida albicans species, two inoculating loops of a standard fungal strain identified as NCP 3179 previously plated on Sabouraud Dextrose Agar which was obtained in collaboration with the Vicente Lemos Laboratory, located in Crato-CE, were inoculated in approximately 60 mL of sterile Brain–Heart Infusion (BHI) liquid medium and also in a solid Sabouraud Dextrose medium, prepared in two sterile plates. This BHI medium was incubated for 24 h at 37 °C, while the Sabouraud medium plates were kept in a humid chamber for approximately 2 weeks. Then, aliquots were prepared from the inoculated medium (Table 1). Thus, 30 samples were prepared, of which 10 were obtained from aliquots taken from the BHI medium and transferred to sterile 15 mL Falcon tubes, referred to as method 1. These samples were centrifuged at 3500 rpm for 5 min to discard the BHI, and then the cells were resuspended in 10 mL of saline solution.

A total of 10 samples were also obtained from each Sabouraud plate using colonies taken directly from this culture medium, referred to as method 2 and method 3, which differ from each other due to adjustments in the nucleic acid extraction protocol. Thus, the maximum number of colonies on the plate was transferred to a sterile Falcon tube containing 10 mL of saline solution, and the suspension was vortexed until it was completely homogenized and separated into aliquots that were transferred to sterile 2.0 mL tubes after removing 20 μL from each sample for cell counting, as shown in Table 1.

Cell counting was performed using the central hemocytometer of the Neubauer Chamber and the total number of cells per mL was estimated by calculating the cells contained in 10 μL (Table 1). Finally, the saline samples stored in 2.0 mL tubes were centrifuged again at 3500 rpm for 5 min to discard the saline solution and stored at −20 °C until extraction.

2.3. Purification of Nucleic Acids

For the extraction of nucleic acids, the method described by Oliveira et al. [15] was associated with the method of Rosa [12], which used SiC with a mesh size of 600 aiming to enhance cell rupture due to its inert and abrasive nature. The reagents used in the extractions were kept refrigerated at 4 °C and stored in an ice bath during the procedure, except for the sodium dodecyl sulfate (SDS) and cetyltrimethylammonium sulfate (CTAB), since they precipitate at low temperatures.

Furthermore, the samples were preserved in an ice bath during the extraction process, except when the protocol recommended room temperature. The agitation, centrifugation, and nucleic acid precipitation time steps described by Oliveira et al. [15] were modified with the aim of reducing the protocol execution time (Table 2) and in the centrifugation steps recommended at 4 °C, the “non-refrigerated” centrifuge rotor was previously kept in the freezer to perform the centrifugations.

Initially, in a 15 mL DNase- and RNase-free Falcon tube containing TE (10 mM Tris, 1 mM EDTA, pH 8.0), SiC (0.35 mg per sample) was added. The mixture was homogenized, and from it, 570 uL were transferred to each sterile and previously labeled 2.0 mL polypropylene tube containing the fungal sample.

Then, 30 μL of 10% SDS was added to each tube and shaken for 10 s manually by inversion. After that, 100 μL of 5 M NaCl/DEPC was added and shaken for 10 s also by inversion. Finally, 100 μL of 10% CTAB solution (CTAB dissolved in 0.7 M NaCl) was added and shaken for 1 min by vortexing; however, the vortexing time was increased to 2 min throughout the protocol in method 3 (Table 2).

After this, 600 uL (1:1) of phenol/chloroform/isoamyl alcohol (24:1) were added and homogenized for 5 min manually by inversion. However, in method 2, this step with phenol/chloroform was performed twice. Then, the samples were mixed by vortexing. Then, the samples were centrifuged at 15,000 rpm for 15 min and the supernatant was transferred to another 2.0 mL microtube, obtaining the final volume for the subsequent addition of 1 volume of (24:1) chloroform/isoamyl alcohol and homogenization by inversion for 5 min at room temperature.

The samples were then centrifuged at 15,000 rpm for 15 min and the supernatant was transferred to another 2.0 mL microtube, obtaining the final volume for the subsequent addition of 0.1 volumes of 3 M sodium acetate (pH 5.0) and two volumes of ice-cold absolute ethanol. The samples were then incubated at −20 °C for 30 min to precipitate the nucleic acids. After this period, a new centrifugation was performed at 15,000 rpm for 15 min.

Finally, 1 mL of 70% ethanol in 1:1000 (v/v) DEPC water was added to the precipitate. Then, everything was centrifuged at 10,000 rpm for 5 min. This step that involved the addition of alcohol, discarding the supernatant, and centrifugation was performed twice. After the last wash, the pellet was dried for approximately 30 min by inversion on absorbent paper. After drying, the nucleic acids were solubilized in 50 uL of water/DEPC and the tubes were left to rest for 15 to 20 min for the subsequent analysis of nucleic acid integrity, as shown in Scheme 1.

2.4. Integrity Analysis

To perform electrophoresis in 1% agarose gel, the step-by-step process was based on the previously consolidated protocols of Sambrook and Russell [16].

2.5. Quantification and Purity of Isolated Nucleic Acids

The evaluation of the purity measurements and quantification of the isolated nucleic acids was obtained in a microvolume spectrophotometer of the Agilent Bio Tek Synergy LX Multimode Reader model Cytation 7 through the Take 3 plate. The test was performed in collaboration with the Molecular Bioprospecting and Alternative Methods Laboratory of URCA, located in Crato-CE. For this purpose, 2 μL of each sample were used. The purity and concentration obtained were A260/A280 (Table 1).

3. Results

The following are the data obtained from the quantification of the samples, the yield, and the purity index obtained by the A260/A280 ratio of methods 1, 2 and 3.

For the extractions in methods 1, 2 and 3, the lowest number of fungal cells obtained and used was 0.025 × 10⁶ CFU by method 1, and 0.595 × 10⁶ CFU was the maximum number of cells obtained by this method (Table 1). Meanwhile, in methods 2 and 3, the highest number of cells obtained, as evidenced in the first samples of these methods, was 0.816 and 0.622 × 10⁶ CFU, respectively, and it is mainly due to the bigger growth of colonies in the solid Sabouraud medium (Table 1).

Regarding the yield obtained by spectrophotometry and number of cells, in method 1, the low concentration of DNA and RNA is directly correlated with the low number of cells. However, in some samples of method 2, especially in sample 1, it is suggested that the low concentration of DNA and RNA is due to an inefficient rupture of the fungal cell wall, since this method presented the largest number of cells (Table 1).

Regarding the yield and purity analysis, in method 3, an “exponential” increase in the extracted nucleic acids was observed in relation to the number of cells from sample 1 to sample 10, along with homogeneous purity among them. Although the yield did not surpass that of method 2 due to the number of cells, method 3 presented superior purity results. This was not observed in the samples from methods 1 and 2 (Table 1), as there were strong indications of the presence of contaminants, such as proteins and phenol.

The results obtained by the 1% agarose gel electrophoresis of the samples from methods 1, 2, and 3 are then displayed for comparison and the subsequent integrity analysis (Figure 1).

When assessing the integrity of the isolated samples using 1% agarose gel electrophoresis, a small pool can be observed, formed by samples 7 to 10 in method 1, 2 to 10 in method 2, and 4 to 10 in method 3. Furthermore, no signs of degradation or deterioration were detected in the gels, particularly regarding RNA. These results indicate that the samples exhibit integrity. Additionally, the pool observed in the samples corresponds to small RNAs, with base pair sizes compatible with 5.8S rRNA and tRNA (Figure 1). Meanwhile, the absence of DNA in the gels is suggestive of a low concentration in the samples, as it is known that the presence of proteins, polysaccharides, phenols or even RNAs does not allow the precise quantification of DNA isolated in the samples by spectrophotometry.

In the table below are the costs for both the nucleic acid extraction and the 1% agarose gel electrophoresis per sample.

4. Discussion

Therefore, the quantification of microorganism cells is relevant not only in fields such as microbiology, public health, and the pharmaceutical industry but also in molecular biology, as a minimum amount of biological sample is required to obtain DNA and/or RNA [17,18,19,20]. This is further reinforced by Silva [21], who states that a sufficient quantity of cells, regardless of the type of sample used, is essential for the stability and proper isolation of genetic material.

Accordingly, this corroborates the findings of Valadares-Inglis and Melo [22], since the success of the isolation is influenced, above all, by the cellular characteristics of the sample—mainly the fungal cellular composition, which differs from animal eukaryotic cells due to the presence of a cell wall that provides resistance and protection [23]. Thus, the partial lysis of the samples, especially of the samples of method 2, is confirmed by Sambrook and Russell [16], when they mention obtaining 5000 to 10,000 ng of RNA per 106 cells depending on the tissue, and evidenced by the maximum RNA yield obtained by the present study, which was 477.6 ng.

Despite the differences between the methods adapted in the present study compared to the standard protocol, according to Beltrão et al. [24] and Oliveira et al. [15], A₂₆₀/A₂₈₀ ratios with values between 1.8 and 2.0 for DNA indicate the presence of a pure sample free from contamination by proteins, carbohydrates, and phenolic compounds. Thus, ratios < 1.8 may suggest protein contamination, while values > 2.0 indicate contamination by reagents, such as phenol. However, in the SiC extraction method performed by Rosa [12], DNA absorbance ratios between 1.55 and 1.78 were considered to be of good quality and with low protein concentrations.

Regarding RNA purity, values close to 2.0 are considered ideal [25]. However, according Petrucelli [26], ratios between 1.8 and 2.0 or even higher values, such as 2.1, are acceptable limits. Therefore, according to the analysis of the spectrophotometry data, method 3 is the one that best encompasses the samples considered pure DNA and RNA simultaneously, as is evident in samples 1, 2, and 3 of this method. For Mello et al. [5], values higher than 30 ng/mL of genetic material, such as those obtained in almost all the samples of methods 2 and 3, are already satisfactory for application in methodologies such as PCR.

The presence of the DNA molecule has not been verified in the gels, because although spectrophotometry detects its presence, according to Viana [27], they may be inflated, above all, by the presence of the RNA molecule. In view of this, additional methods are needed to really evaluate the presence of this molecule, such as amplification by PCR, given that for Barea [28], in some extraction methods the spectrophotometric results are not reliable parameters regarding its quantification, because even when the measurements did not reveal its presence in the sample, there was, however, its amplification.

Therefore, the results obtained in electrophoresis were promising. In addition to corroborating the yield and purity analyses obtained by spectrophotometry in the present study, it is also important to consider the specific characteristics of RNA, which, according to Devlin [29], make this molecule very unstable and easily degradable. For this reason, Dettogni and Louro [30] state that RNA is so fragile that it deserves special attention during the extraction process, and methods optimized specifically for its protection due to the difficulty of laboratories in isolating this genetic material in some biological samples.

Regarding the cost–benefit analysis of using a manual protocol for joint DNA/RNA extraction and integrity assessment, it is evident that the adapted protocol offers excellent cost-efficiency, at approximately BRL 0.937 per sample, and allows multiple extractions to be performed (Table 3). In contrast, commercial kits, although effective in isolating high-quality nucleic acids, have the disadvantage of high cost per sample, around BRL 6.00 according to Araújo et al. [31]. This aligns with the findings of Amaral et al. [32], who, when comparing phenol/chloroform extraction with other methodologies, including a commercial kit, classified it as the most efficient method due to its ability to provide high-quality and well-concentrated genetic material at a low cost, with a relatively fast processing time (2 h and 30 min) compared to other methods (8 h and 30 min). The present study confirms these findings, demonstrating that the adapted protocol, derived from a standard protocol of 6 h and 50 min, can be completed in a maximum of 2 h and 12 min.

In terms of reproducibility, the adapted protocols follow the same steps as standard protocol, although they are executed on a shorter time scale. As such, the standard protocol is not the best option for small- and medium-sized laboratories, as it becomes time-consuming for higher workloads typical of these environments. Regarding the reproducibility of the adapted methods with silicon carbide, they isolated genetic material with excellent quality and high yield through a fast, low-cost approach similar to the unpublished study by Santos [33] who also obtained similar and promising results in the isolation of bacterial genetic material using the same SiC-based adapted methodology.

5. Conclusions

The protocol adapted in these methods proved to be highly promising for obtaining pure and complete samples, suitable for use in various molecular tools, in less time. As a result, it offers a cheaper, faster, and simpler method, with excellent results achieved using minimal amounts of biological sample. This makes it an accessible methodology applicable to various laboratory infrastructures, ranging from research in small laboratories to routine use in larger facilities.

Author Contributions

E.C.V.d.F., M.R.V.L. and D.L.d.S.J. wrote the manuscript and compiled the data; F.A.d.S., A.C.F.A., P.R.F. and T.T.A.Y. carried out the microbiological and molecular biology experiments; M.K.d.N.S.L. coordinated the project and carried out the tests; I.R.A.d.M. and H.D.M.C. took part in revising and finalizing the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

All data will be available after a reasonable request to the corresponding authors.

Conflicts of Interest

The authors declare no conflicts of interest.

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Scheme 1. Nucleic acid purification and precipitation process. Source: Prepared by the author on the Biorender platform.

Figure 1. Nucleic acid integrity analysis in 1% agarose gel. M: DNA Ladder 1 kb Plus—exACTGene Fischer BioReagent; (A) samples 1 to 10, method 1; (B) method 2; (C) method 3.

Table 1. Concentration and purity of total nucleic acids isolated from Candida albicans.

				Performance (ng/μL)		Purity
				Performance (ng/μL)		(A260/A280)
Cultivation	Sample	Average Aliquot (mL)	No. cells/mL (× 10⁶ CFU)	DNA	RNA	DNA	RNA
Method 1 (BHI)	1	0.1	0.025	-	-	-	-
	2	0.5	0.075	-	-	-	-
	3	1.0	0.115	-	-	-	-
	4	1.5	0.115	-	-	-	-
	5	2.0	0.160	24.8	-	1.3	-
	6	2.5	0.260	-	-	-	-
	7	3.0	0.270	59.3	26.0	1.4	1.2
	8	3.5	0.295	72.4	36.1	1.4	1.2
	9	4.0	0.405	57.7	25.6	1.3	1.13
	10	4.5	0.595	61.9	31.1	1.4	1.1
Method 2 (Sabouraud)	1	0.2	0.816	7.9	50.7	1.0	1.3
	2	0.4	1.632	86.9	-	1.4	-
	3	0.6	2.448	126.1	84.0	1.5	1.4
	4	0.8	3.264	102.3	61.9	1.6	1.4
	5	1.0	4.080	93.9	49.0	1.6	1.4
	6	1,2	4.896	263.7	231.4	2.1	2.2
	7	1.4	5.712	325.2	288.2	2.2	2.2
	8	1.6	6.528	165.9	145.5	2.1	2.1
	9	1.8	7.344	354.9	302.8	2.1	2.2
	10	2.0	8.160	569.8	477.6	2.2	2.2
Method 3 (Sabouraud)	1	0.2	0.622	35.2	29.3	2.0	2.0
	2	0.4	1.244	47.5	40.4	1.8	1.7
	3	0.6	1.867	49.8	42.4	2.0	2.0
	4	0.8	2.489	97.7	83.4	2.1	2.1
	5	1.0	3.112	126.5	108.1	2.1	2.1
	6	1,2	3.734	202.0	171.9	2.2	2.2
	7	1.4	4.356	209.6	177.1	2.2	2.2
	8	1.6	4.979	228.9	190.4	2.2	2.2
	9	1.8	5.601	220.2	185.7	2.2	2.2
	10	2.0	6.224	384.2	312.6	2.2	2.2

Source: Prepared by the author.

Table 2. Modified stages versus the method of Oliveira et al. [15].

		Adapted Method
Steps	Oliveira et al. [15]	Methods 1 and 2	Method 3
Cell sedimentation	2 min/10,000× g (4th)	5 min/3500 rpm *	5 min/3500 rpm *
Homogenized by inversion	10 s	10 s	10 s
Vortex homogenization	1 h (200 rpm)	1 min *	2 min *
Homogenized by inversion	10 s	10 s	10 s
Vortex homogenization	1 h (200 rpm)	1 min *	2 min *
Homogenized by inversion	10 min	5 min *	5 min *
Centrifugation	15 min/10,000× g (4th)	15 min/15,000 rpm *	15 min./15,000 rpm *
Homogenized by inversion	10 min	5 min *	5 min *
Centrifugation (4th)	10 min/10,000× g (4th)	15 min/15,000 rpm *	15 min/15,000 rpm *
Precipitation	3 h at −20 °C	30 min at −20 °C *	30 min at −20 °C *
Centrifugation	10 min/10,000× g (4th)	15 min/15,000 rpm *	15 min/15,000 rpm *
Centrifugation	5 min/7000× g (4th)	5 min/10,000 rpm *	5 min/10,000 rpm *
Drying	30 min	30 min	30 min
Solubilized in 50 μL of water	-------	-------	------
Execution time	06 h 50 min 06 s	02 h 10 min 50 s	02 h 12 min 50 s

* modified steps. Source: Prepared by the author.

Table 3. Costs per sample for performing joint extraction of genetic material and 1% agarose gel electrophoresis.

DNA/RNA Extraction—Mechanical Method (Silicon Carbide)			By Sample
Reagents	Grams/mL Per Sample (0.5 mL)	Mark	BRL
Tris (hydroxymethyl)aminomethane	0.06056 gold	LGC 250 g	0.094
Disodium salt EDTA (2H₂O) PA	0.01861 gold	Neon 500 g	0.0041
Sodium chloride PA	0.01957 gold	Neon 500 g	0.00087
Cetyltrimethylammonium bromideSHOVEL	0.001 gold	Dynamic 100 g	0.0009
Sodium Dodecyl Sulfate (SDS)	0.003 gold	Dynamic 500 g	0.00096
Phenol 90%	0.6 mL	Dynamics 1 L	0.064
Chloroform PA	1.2 mL	Dynamics 1 L	0.216
Isoamyl alcohol PA	0.024 mL	Dynamics 1 L	0.0036
Sodium acetate PA	0.0073 gold	Dynamic 500 g	0.0008
Ethanol PA	1.8 mL	Dynamics 1 L	0.090
Diethylpyrocarbonate (DEPC)	0.015 mL	Sigma-Aldrich-25 mL	0.462
By sample	-	-	0.937
DNA/RNA electrophoresis
Reagents	TAE 1X (300 mL)	Mark	BRL
Tris PA	1.452 g	LGC 250 g	2.26
EDTA PA	0.124 gold	Neon 500 g	0.021
Acetic acid	0.34 mL	Dynamics 1 L	0.061
Diethylpyrocarbonate	0.3 mL	Sigma-Aldrich 25 mL	9.24
Agarose PA	0.30 gold	KASVI 100 g	1.476
IntercalatingBlue Green Loading Buffer Dye I (0.6 mL)	0.0006 mL (0.6 μL)		0.17
DNA Ladder 1 kb plus (0.5 mL)	0.0008 mL (8 μL)	LGC Biotechnology	1.696
Total (per gel)	-	-	14.92

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MDPI and ACS Style

Freitas, E.C.V.d.; Santos, F.A.d.; Lopes, M.R.V.; Sousa Júnior, D.L.d.; Yafawi, T.T.A.; Araújo, A.C.F.; Freitas, P.R.; Menezes, I.R.A.d.; Coutinho, H.D.M.; Leandro, M.K.d.N.S. Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells. DNA 2024, 4, 417-426. https://doi.org/10.3390/dna4040029

AMA Style

Freitas ECVd, Santos FAd, Lopes MRV, Sousa Júnior DLd, Yafawi TTA, Araújo ACF, Freitas PR, Menezes IRAd, Coutinho HDM, Leandro MKdNS. Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells. DNA. 2024; 4(4):417-426. https://doi.org/10.3390/dna4040029

Chicago/Turabian Style

Freitas, Elizabeth Cristina Vieira de, Francisca Alves dos Santos, Maria Raíssa Vieira Lopes, Dárcio Luiz de Sousa Júnior, Tássia Thaís Al Yafawi, Ana Carolina Ferreira Araújo, Priscilla Ramos Freitas, Irwin Rose Alencar de Menezes, Henrique Douglas Melo Coutinho, and Maria Karollyna do Nascimento Silva Leandro. 2024. "Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells" DNA 4, no. 4: 417-426. https://doi.org/10.3390/dna4040029

APA Style

Freitas, E. C. V. d., Santos, F. A. d., Lopes, M. R. V., Sousa Júnior, D. L. d., Yafawi, T. T. A., Araújo, A. C. F., Freitas, P. R., Menezes, I. R. A. d., Coutinho, H. D. M., & Leandro, M. K. d. N. S. (2024). Co-Extraction of DNA and RNA from Candida albicans Using a Chemical Method in Conjunction with Silicon Carbide with Few Cells. DNA, 4(4), 417-426. https://doi.org/10.3390/dna4040029

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