Multiple tube technique

Multiple tube technique No more than 10% portions (sample tubes) positive ... 

Quantitative tests. In some situations, actual enumeration of the number of organisms present may be necessary. This is the case, for example, for limitations imposed on a discharge permit. Generally, two methods are used to enumerate conforms the membrane-filter technique and the multiple-tube technique. 

The other method of enumerating coliforms is through the use of the multiple-tube technique. This method is statistical in nature and the result is reported as the most probable number (MPN) of organisms. Hence, the other name of this method is the MPN technique. This technique is an extension of the qualitative techniques of presumptive, confirmed, and completed tests. In other words, MPN results can be a presumptive, confirmed, and completed MPNs. The number of tubes liberating gases is counted from each of the set of hve tubes. This information is then used to compute the most probable number of organisms in the sample per 100 mL. 

Most Probable Number Method (Multiple Tube Technique).112... 

One standard test used to determine the presence of the coliform group is called the multiple-tube fermentation technique (sometimes called the presumptive test). If this test indicates the presence of these bacteria, then a confirmed test must be done. If only negative colonies or no colonies develop during this test, it is considered negative otherwise, a completed test must be undertaken. Positive results obtained in the completed test are evidence for the presence of coliform bacteria. Testing methods have been given by the APHA, and the detailed procedures contained therein should be consulted (20). 

The water analysis is incomplete unless the number of coliform bacteria present is determined as well. A multiple-tube fermentation technique can be used to enumerate positive presumptive, confirmed, and fecal coliform tests. Results of the tests are expressed in terms of the most probable number (MPN). That is, the count is based on a statistical analysis of sets of tubes in a series of serial dilutions. MPN is related to a sample volume of 100 ml. Thus, an MPN of 10 means 10 coliforms per 100 ml of water. 

Zarowin (68) has made use of a multiple-sampling technique in the measurement of decay times. This method uses a periodically pulsed- or chopped-excitation source and a continuously operating photomultiplier detector. The fluorescent signal is displayed on an oscilloscope. The response of the photomultiplier tube must be fast enough to resolve individual photoelectron pulses, and the time density of pulses is then proportional to the light intensity. 

Multiple-tube or MPN technique—A method of estimating the concentration of conforms using the Poisson distribution. 

Multiple tube methods are very sensitive to small numbers of micro-organisms due to the powerful amplification of these techniques. However, the methods are not very accurate statistically, and have considerable errors associated with them. In addition, the multiple dilutions needed are very demanding on technician time, sterile glassware and media. Tube methods may be run in conjunction with plate counts. 

Four methods may be used to detect and enumerate total coliforms, fecal coliforms, and E. coli in water samples. These are grouped as (a) multiple tube fermentation technique also known as the MPN technique, (b) membrane filtration technique, (c) presence/absence test, and (d) use of the enzymatic substrates test also known as the chromogenic-fluorogenic substrate test. 

Gonadotrophins are also used in assisted reproduction procedures. Here the aim is to administer therapeutic doses of FSH that exceed individual follicular FSH threshold requirements, thus stimulating multiple follicular growth. This, in turn, facilitates harvest of multiple eggs, which are then available for in vitro fertilization. This technique is often employed when a woman has a blocked fallopian tube or some other impediment to normal fertilization of the egg by a sperm cell. After treatment, the resultant eggs are collected, incubated in vitro with her partner s sperm, incubated in culture media until the embryonic blastocyst is formed, and then implanted into the mother s uterus. 

The advantage of trimethylsilyl (TMS) derivatives lies in the simplicity of the derivatization procedure, which is carried out by the addition of N,0-bis(trimethylsilyl)trifluoroacetamide (BSTFA) in acetonitrile and heating for approximately 2 h at 150 °C under anhydrous conditions in a sealed tube. However, there may be problems owing to the formation of multiple derivatives of each amino acid. Another technique involves the formation of n-butyl esters of the amino acids and their subsequent trimethylsilylation by a similar procedure. The n-butyl esters are formed by heating the amino acids for 15 min in n-butanol and HC1 and these are then converted to the A-TMS-n-butyl ester derivatives. A-acyl amino acid alkyl esters are commonly used. Acetylation of the butyl, methyl or propyl esters of amino acids,... 

This ground-breaking technique for making multiple copies of DNA in a test tube was invented in the 1980s by Katy Mullis, a biochemist. Mullis revolutionary technique earned him a Nobel Prize in Chemistry in 1993. 

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