Lab 6: Extraction of Plasmid DNA using GF-1 Plasmid DNA extraction kit


Introduction:


The GF-1 Plasmid DNA extraction kit is a device for extraction of plasmid DNA in a fast and efficient purification withoud the need for precipitation or organic extraction from lysates. This kit uses alkaline lysis-SDS method to lyse cells and release plasmid DNA.


plasmid DNA extraction kit



This device use a special-treated glass filter membrane fixed into a column to efficiently bind DNA in the presence of high salt. Special byffers provided in the kit are optimized to enhance binding of DNA.


Features:
- Yields up to 20µg of DNA
- Multiple samples can be processed rapidly in less     than 30  minutes
- No organic-based extraction required
- Highly pure plasmid DNA ready to use for routine   molecular biology applications such as restriction enzyme digestion, PCR, DNA sequencing, ligation, transformation, etc. 


Procedure: 





instruction from our lab instructor 

Result:








Discussion:

GF- 1 Plasmid DNA extraction kid is designed for rapid and efficient purification of high copy and low copy plasmid DNA withoud need for precipitation organic extraction. Plasmid are often used to purify a specific sequence, since they can easily be purified away from the rest of the renomw. For their uses as a vector, and for molecular cloning, plasmid often need to be isolated.There are several methods to isolate plasmid DNA from bacteria, the archetypes of which are the miniprep and the maxiprep. The former can be used to find out whether the plasmid is correct in any of several bacterial clones faster. The yield is a small amount of impure plasmid DNA, which is sufficient for analysis by restriction digest and for some cloning techniques.In the latter, much larger volumes of bacterial suspension are grown from which a maxi-prep can be performed. Essentially this is a scaled-up miniprep followed by additional purification. This results in relatively large amounts (several micrograms) of very pure plasmid DNA.


column-assembled collection tube

collection tube contain culture



Conclusion:

This report has identified that GF-1 Plasmid DNA extraction kit is the best,fast and efficient way to extract plasmid from DNA. This kit does not need precipitation or organic extraction from lysates and processes multiple samples in less than 30 minutes. Besides, GF-1 Plasmid DNA suitables for many molecular biology applications such as restriction enzyme digestion, PCR, DNA sequencing, ligation, and transformation.


References:

http://faculty.plattsburgh.edu/donald.slish/Extraction.html
http://en.wikipedia.org/wiki/File:Conjugation.svg
http://en.wikipedia.org/wiki/Plasmid
http://www.ncbi.nlm.nih.gov/pubmed/20626237
01:19 | Category: | 0 comments

Lab 5: Determination of Antimicrobial Effects of Microbial Extracts.


Introduction:

Multiwavelength ultraviolet/visible (Uv-vis) spectroscopy is a versatile,
quantitative, rapid, and reliable analytical tool that has immediate applications as a biosensor for the detection, identification, and enumeration of microorganisms and cells. The sample information contained in a spectrum includes cell size, chemical composition, shape, and information on their internal structure.  This information is
obtained from the spectroscopic analysis of  a sample measured over a broad range of wavelengths (200-900 nm) and/or with the scattered light measured at one or many different angles. The potential to extract  large amounts of information from a single multiwavelength measurement makes Uv-vis spectroscopy a powerful characterization
tool. In addition, process Uv-vis spectrometers and miniaturized systems, make this technique readily available for real-time in situ monitoring of biological, and environmental processes.

Bacteriocins comprise a large and diverse group of ribosomally synthesised antimicrobial proteins or peptides. Although bacteriocins can be found in numerous Gram-positive and Gram-negative bacteria, those produced by lactic acid bacteria (LAB) have receive special attention in recent years due to their poential application in the food industry as natural biopreservatives. Different classes of LAB bacteriocin have been identified on the basis of biochemical and genetic characterization. These bacteriocins have been reported to inhibit the growth of Listeria monocytogenes, Staphylococcus aureus, Enterococcus faecalis and Clostridium tyrobutyricum.

preparation of sample

prepared sample




Result:






Discussion:

Multiwavelength Uv-vis spectra of microorganisms and cell suspensions contain quantitative information on their properties such as number, size, shape, chemical composition, and internal structure. These properties are essential for the identification and classification of cells.  The complexity of microorganisms in terms of their chemical
composition and internal structure make the interpretation of their spectral signature a difficult task. In this lab report  proposed for the interpretation of the multiwavelength spectra of microorganisms.
The optical properties as functions of wavelength, and available literature data on the size and chemical composition of E. coli cells and S. aureus have been used to explore the sensitivity of the calculated spectra to the model parameters. It is shown that the proposed model can reproduce the features of experimentally measured spectra. The sensitivity of the spectra to the model parameters suggests that the proposed model can be used for the quantitative deconvolution of the
Uv-vis spectra in terms of critical information necessary for the detection and identification of microorganisms.  


An anti-microbial is a substance that kills or inhibits the growth of microorganisms such as bacteria, fungi, or protozoans. Antimicrobial drugs either kill microbes (microbiocidal) or prevent the growth of microbes (microbiostatic). Disinfectants are antimicrobial substances used on non-living objects or outside the body.
A drug used to treat a microbial infection. "Antimicrobial" is a general term that refers to a group of drugs that includes antibiotics, antifungals, antiprotozoals, and antivirals.

Conclusion:

A model for the interpretation of the multiwavelength Uv-vis spectra of microorganisms and cells has been formulated. The proposed interpretation model is based on light scattering theory, spectral deconvolution techniques, and on the approximation of the
frequency dependent optical properties of the basic constituents of living organisms.


Reference:

http://www.sciencedirect.com/science?







23:29 | Category: | 0 comments

Lab 4: Source of Contamination and Infection



Introduction:

A microorganism or microbe is an organism that is unicellular or lives in a colony of cellular organisms. Microorganisms contamination is a situation which occurs when microorganisms end up in a location where they are not supposed to be. It is often used to refer to contamination of food by bacteria which can cause disease, but microorganisms contamination can also occur in other settings. This situation is not desirable, because it can pose a health threat and cause other problems. As microorganisms, particularly bacteria, are found practically everywhere, this means in most cases the reduction of harmful microorganisms to acceptable levels. However, in some cases it is required that an object or substance be completely sterile, i.e. devoid of all living entities and viruses.
Although a number of microorganisms are present in air, it doesn't have an indigenous flora. Air is not a natural environment for microorganisms as it doesn't contain enough moisture and nutrients to support their growth and reproduction.
In food preparation microorganisms are reduced by preservation methods (such as the addition of vinegar), clean utensils used in preparation, short storage periods or by cool temperatures. If complete sterility is needed, the two most common methods are irradiation and the use of an autoclave, which resembles a pressure cooker. There are no conditions where all microorganisms would grow, and therefore often several different methods are needed. 




Preparation of the sample



Result:



All petri dish after incubate at 37°C for 48 hours

A petri dish contain wash water


collecting sample from ear



Disscusion:


Air currents may bring the microorganisms from plant or animal surfaces into air. These organisms may be either commensals or plant or animal pathogens. However, the transmission of animal diseases is not usually important in outside air.

The main source of airborne microorganisms is human beings. Their surface flora may be shed at times and may be disseminated into the air. Similarly, the commensal as well as pathogenic flora of the upper respiratory tract and the mouth are constantly discharged into the air by activities like coughing, sneezing, talking and laughing. The microorganisms are discharged out in three different forms which are grouped on the basis of their relative size and moisture content. They are droplets, droplet nuclei and infectious dust.
Everyone knows that flu viruses and cold germs can congregate on hands. Also found on the hands are the bacteria that cause gastrointestinal problems. One of the most common types of bacteria known to man is staph. Because in recent years staph is becoming increasingly resistant to antibiotics which can prove deadly, washing your hands is more than good hygiene, it can save your life.

The respiratory system is open to airborne microbes and to outside pollution.It is not surprising that respiratory diseases occur, in spite of the body's defenses. Some respiratory disorders are relatively mild and, unfortunately, very familiar. We all experience the excess mucus, coughing, and sneezing ofthe common cold from time to time. The common cold is an example of rhinitis, an inflammation of the epithelium lining the nose and nasal cavity. Viruses, bacteria, and allergens are among the causes of rhinitis. Other respiratory disorders include laryngitis, pneumonia, bronchitis, chronic obstructive pulmonary disease, and lung cancer.

The cough reflex is a vital part of the body's defence mechanisms. Normally, the lungs and the lower respiratory passages are sterile. If dust or dirt get into the lungs, they could become a breeding ground for bacteria and cause pneumonia or infection in the breathing tubes. When a person coughs, there is a short intake of breath and the larynx (the voice box) closes momentarily. The resulting blast of air comes out at high speed, scrubbing and clearing the airway of dust, dirt or excessive secretions.

Conclusion:


  • Bacteria are everywhere and can spread from surface to surface, person to person, food to food, and person to food. Harmful bacteria can be controlled by practicing the 4 Cs of food safety. To prevent the spread of harmful bacteria, proper cleaning of both hands and surfaces is especially important.  The good thing is that not all bacteria are harmful; most bacteria are beneficial to us.
  • When designing experiments, you should always use safe techniques when working with bacteria. Also, it's important to have a control plate. In this experiment, you also learned that different strains of bacteria can be identified through colony morphology.
References:

http://www.microbiologyprocedure.com/air-microbiology/sources-of-microorganisms-in-air.htm
http://en.wikipedia.org/wiki/Microorganism
http://www.ehow.com/hand-bacteria/
http://www.netdoctor.co.uk/health_advice/facts/cough.htm
http://www.wisegeek.com/what-is-bacterial-contamination.htm    
http://www.ageds.iastate.edu/meat/topic1/bacteriaeverywhere.htm
10:31 | Category: | 0 comments

our COMMITMENT!!!!!!!