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Register a new userGenomic DNA extraction from small volume specimens using different methods and assessment its concentration and purification
استخلاص الDNA المورثي من العينات صغيرة الحجم بطرق مختلفة و تقييم تركيزه و نقاوته
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تم في هذه الدراسة استخلاص DNA من شرائح التجري المجهري عن الإصابة بداء الليشمانية. استخدمت ثلاث طرائق لاستخلاص DNA الأولى باستخدام محاليل ملحية جاهزة Promega Kit و الثانية تقليدية و الثالثة تعتمد استخدام أعمدة مزودة بأغشية من مادة سيليكاجيل طريقة Qia gen.
DNA was extraction from scanning microscopic slides to identification leishmaniases. Three methods used to DNA extraction, first one using ready extraction solvent (Promega kit), second depended on phenol-chloroform-iso propanol , whereas third one depended on columns with silica gel membranes (Qia gen kit). The results showed that (Promega kit) method didn’t gave high concentration of DNA. (Qia gen kit) method was the best for DNA concentration and purity
References used
Alger J, Martha C Acosta, Caterin Lozano, Carmelita Velasquez, Luz Angela Labrada.(1996) Stained Smears as a Source of DNA. Mem Inst Oswaldo Cruz, Rio de Janeiro, Vol. 91(5): 589-591
Al-Nahhas Samar and Kaldas Rania . (2013): Characterization of Leishmania Species Isolated from Cutaneous Human Samples from Central Region of Syria by RFLP Analysis. ISRN Parasitology. Article ID 308726, 5 pages
Desjeux P 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27: 305-318
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The aim of this research, is to use Liquid–liquid extraction to separate
phenolic compounds from olive mill wastewater resulted from three-phase
centerfuge extraction (OMWW). Four solvents were used: Methyl iso butyl
ketone (MIBK), Ethyl acetate (
EtOAc), Octanol-1 (C8H18O), and Noctanoylpyrrolidine
(OPOD) which was synthesized in our lab. The effects of
pH and Extraction stages on the efficiency of total phenolic compound
extracted from OMWW were determined by Folin - Ciocalteu
spectrophotometric method. Ferulic acid (FA) and Caffeic acid (CA) were
determined in extractants by HPLC. The COD value in water phase of the four
extracts was measured together with the reducing power and radicalscavenging
effects for the extracts. We compared the antioxidant activity of
MIBK, OPOD extracts, with ferulic acid (FA), caffeic acid (CA), ascorbic acid
standards and rosemary.
The results showed that MIBK had the highest extraction efficiency and
gave the maximum quantity of phenols from OMWW, than OPOD, C8H18O,
EtOAc. The COD value was decreased about %94.5 by MIBK, %83.7 by
OPOD,%81 by C8H18O, and%72 by EtOAc. The phenols extracted by MIBK
could be added to eatable oils as natural antioxidants, because they were
superiority on rosemary, and nearly to caffeic acid (CA) standard.
The study included the selection of the best methods to extract the enzyme
among nine methods. The protein content was concentrated and precipitated
by cold acetone among other five methods of concentration (partial
purification). The purification
stages were achieved by using ion exchange
column chromatography (DEAE – Sephadex A 50 column). Followed by gel
filtration chromatography using sephacryl S-300. The active parts were
lyophlizated (free drying) to obtain Lipoxygenase with 43.18% yield and 8.16
folds of purification and specific activity of 1162.9 unit / mg.
The purity of enzyme was confirmed by poly acryl amide gel
electrophoresis under non denaturing conditions, with the appearance a single
band .
The experiment was carried out at the end of April during 2013 In Abi-
Jarash farm, Faculty of Agriculture, Damascus University in order to
determine the influence of organic fertilization on the growth and yield of Snap
bean. The snap bean ”Tema”
Phaseolus Varity, and organic fertilizer” Humax”
with two methods (root application and foliar spry) and two concentrations (2
and 4 g/L) were used. The organic fertilizer was used after two weeks of
germination and then two weeks between treatment and another and the
experiment was laid according to simple randomized block design. Results
showed that organic fertilizer caused an increase of plants growth and yield,
and the foliar spray method showed significant superiority (p<0.05) to some of
the studied parameter such as plant dry weight (6.89 g), leaf area (4595.44 cm2),
leaf area index (2.55), number of clusters (15.3 cluster/ plant), number of pods
(29.6 pod/plant), plant yield (156.9 g/plant), compared to the root applied
method. Results also showed no significant effect for the fertilizer
concentration.
Several protocols for DNA extraction from leaves of pepper Capsicum annuum
L. were evaluated to detect Begomoviruses. The extraction
methods comparison were based on DNA quantity and quality. The
DNA concentration absorbance for the evaluation of DNA concentration and contamination
were measured using spectrophotometer.
β-galactosidase enzyme was isolated from the new born goat brain by nine
methods, It was found that the sodium acetate 0.2 Mole/Liter +0.2Mole/Liter
NaCl PH5 method have given the highest specific activity of crude enzyme in
comparison with the ot
her methods. Also, this enzyme was purified by using
four methods, the second one (cold acetone) was the butter. As a result the
purification fold was about 135.46 times and the yield about 77.14% by using
Sephacryl S200 (second step). This enzyme is 187.437 KDa as a molecular
weight.
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