Showing 4 results for Extraction
A Alavi, Sh Moradi, N Mirkheshti, A Ghadiri, F Hadizadeh,
Volume 1, Issue 1 (4-2007)
Abstract
Abstract Background and objectives: Hemin is a porphyrin compound derived from hemoglobin, the precursor of other porphyrin hemoglobin derivatives and the raw material of Hematin. Since hemin is widely used in medicine, we decided primarily to synthesize this substance in Laboratory and to determine the best way of hemin extraction from untransfused and expired blood units. Materials and Methods: In the first method, Glacial acetic acid and sodium chloride were added to citrated blood and hemin crystals were extracted by means of cooling. Finally, the obtained product, by visible spectrophotometer and Infrared Spectrophotometer, was compared to standard samples. Fur thermore, citrated blood, citrated blood hemolysed by distilled water and citrated blood washed by normal saline were used comparatively as a raw material to produce Hemin. The second method was performed by adding Strontium, acetic acid and acetone to blood samples and then after precipitating Hemin crystals they were washed and dried with acetone. Results: The presence of functional groups in Hemin samples, analyzed by infrared Spectrophotometer, indicates the production of this compound. The results of visible Spectrophotometer in comparison with control samples and the results of samples weighting demonstrates high efficiency of extraction stages and the purity of obtained compound. Conclusion: The use of intact citrated blood produces more Hemin than the other kind of Citrated blood samples. Moreover, acetic acid with citrated blood, without any processing on blood, is the best way for Hemin production. Key words: strontium, Hemin, Blood, acetic acid, extraction
H Davoodi, S R Hashemi, H F Seow, M Ghorbani,
Volume 3, Issue 2 (10-2009)
Abstract
Abstract Background and objectives: Paraffin-embedded tissues and clinical samples are a valuable resource for molecular genetic studies, but the extraction of high-quality genomic DNA from this tissues is still a problematic issue. In the Present study, the efficiency of two DNA extraction protocols, a commercial kit and a traditional method based on heating and K Proteinase was compared. Material and Methods: Fifty paraffin-embedded blocks of colon cancer tissues (more than 5 years old) were used to compare two methods of DNA extraction. DNA was extracted by traditional method using heat and commercial DNA extraction (Qiagen kit) method. Then the purity and concentration of extracted DNA were measured by Spectrophotometer. Two sequences of TLR4 “The most important receptors in innate immunity” were amplified by polymerase chain reaction. SH-1 ‘188bp’ and SH-2 ‘124bp’ were amplified and then the products were separated on a 2% agarose gel. Results: The results show that the yield of DNA by traditional method (297 mg/ml) is significantly (p<0.01) higher than Commercial kit (176mg/ ml). But traditional method has the lower OD ratio (1.2) Compared to Commercial method. The Amplification of the TLR4 gene sequences is more successful by the traditional method (p<0.01) compared with commercial method. The length of the sequence affects on the results of PCR in that short sequence is amplified more successful compared to the long sequence. Conclusion: The traditional method is more successful in PCR amplification and also simple and cheap. Therefore, we recommend using this method for DNA extraction taken from the paraffin-embedded blocks with more than 5 years old and selecting shorter sequence for better amplification in PCR. Key words: DNA Extraction, paraffin embedded tissue, PCR
Mousazade Moghadam M, Babavalian H, Mirnejad R, Shakeri F,
Volume 6, Issue 1 (4-2012)
Abstract
Abstract
Background and objectives: Genomic DNA extraction of bacterial cells is of processes performed normally in most biological laboratories therefore, various methods have been offered, manually and kit, which may be time consuming and costly. In this paper, genomic DNA extraction of Staphylococcus aureus was investigated using some laundry detergent brands available in Iran to achieve a rapid and cost effective method.
Material and Methods: five-enzyme Taj brand, three-enzyme Saftlan brand ,and Darya and Pak brands without enzyme were used in the concentrations of 10, 20, 40, 80 mg/L. Afterwards, in order to evaluate the efficiency of extracted DNA in downstream processing, PCR test was performed for femA gene in the genome of Staphylococcus aureus.
Results: DNA extraction using different concentrations of the brands show that extracted DNA using 40 mg/L Saftlan and Taj brand powders have the best results according concentration (µg/ml) and purity (A260/A280) parameters. These parameters are 387.5 1.88 (Taj), 254.1 2.80 (Softlan), 396.6 1.95 (Manual) and 423.3 2.2 (Kit), respectively. Afterward, the PCR test results by show that DNA extraction using laundry detergents has no effect on its efficiency in order to be used in downstream processes.
Conclusion: These results indicate that the proper concentrations of laundry detergents can be used to extract genomic DNA with similar efficiency to kit and manual extraction methods.
Key words: Bacterial genome, DNA extraction, laundry powder, PCR, Staphylococcus aureus
Fereshteh Keyghobadi, Nader Bahramifar, Elahe Gharekhani, Seyyedeh Marzieh Kia,
Volume 13, Issue 5 (9-2019)
Abstract
ABSTRACT
Background and Objectives: In this study, nanosilica modified with HS-SiO2 thiol groups was utilized as adsorbent for solid phase extraction, as a fast and reliable method of preconcentration and separation of very small quantities of selenium ions from water and blood samples.
Methods: The samples included four natural water samples and one biological sample (blood serum) prepared in volumes of 25, 100, 200, 300, 400 and 500 ml. The samples were analyzed by solid phase microextraction, using thiolated-nanosilica (as adsorbent), ultraviolet-visible spectrophotometry and atomic absorption spectroscopy.
Results: Optimized conditions for preconcentration of a 25 ml 0.2 mg/l selenium solution were pH 5, 40 mg of adsorbent, sample-adsorbent mixing time of 15 minutes and 5 ml of 2N sulfuric acid as detergent. The volume limit and concentration factor were 400 and 80, respectively. Limit of detection and relative standard deviation of the method were 0.46 μg/l and 0.9%, respectively.
Conclusion: This study is the first to successfully utilize thiolated nanosilica for measuring low selenium levels. Thiolation of the absorbent increases selenium adsorption by thiolated-silica compared to SiO2.
Keywords: Solid phase extraction, Selenium, Preconcentration, Nano, UV-visible spectrophotometry.
