Summary of Methods to Identify, Enumerate and Characterize Bacteria and Fungi In Soil

-
Automated rRNA intergenic spacer analysis (ARISA) was used to characterise bacterial (B-ARISA) and fungal (F-ARISA) communities from different soil types. The 16S-23S intergenic spacer region from the bacterial rRNA operon was amplified from total soil community DNA for B-ARISA. Similarly, the two internal transcribed spacers and the 5.8S rRNA gene (ITS1-5.8S-ITS2) from the fungal rRNA operon were amplified from total soil community DNA for F-ARISA. Universal fluorescence-labeled primers were used for the PCRs, and fragments of between 200 and 1,200 bp were resolved on denaturing polyacrylamide gels by use of an automated sequencer with laser detection. Methodological (DNA extraction and PCR amplification) and biological (inter- and intrasite) variations were evaluated by comparing the number and intensity of peaks (bands) between electrophoregrams (profiles) and by multivariate analysis. Our results showed that ARISA is a high-resolution, highly reproducible technique and is a robust method for discriminating between microbial communities. To evaluate the potential biases in community description provided by ARISA, we also examined databases on length distribution of ribosomal intergenic spacers among bacteria (L. Ranjard, E. Brothier, and S. Nazaret, Appl. Environ. Microbiol. 66:5334–5339, 2000) and fungi.
Source Link : http://www.ncbi.nlm.nih.gov/pmc/articles/PMC93193/

Comments

Post a Comment

Popular posts from this blog

Prepare 6X Laemmli buffer

-
Laemmli is a sample buffer to use in western blot. 6X sample buffer is added to each protein sample and is boiled or heated for 5-10 minutes. Recipe to prepare 10 ml: - 1.2gr SDS (sodium dodecyl sulfate) - 6mg bromophenol blue - 4.7ml glycerol - 1.2ml Tris 0.5M pH6.8 - 2.1ml dwater warm it a little bit and shake it till everything is dissolved. It takes time. - add 0.93gr DTT. Wait until it is completely dissolved. Aliquote and keep frozen at-20ºC.
Read more

0.1 M Triethanolamine

-
0.1 M TEA: TEA = Triethanolamine You will dilute TEA 1:10 in Millipore water: Pour 50 mL TEA into a 50 mL conical tube (TEA is very viscuous, so best to measure it in something that is disposable). Pour TEA from conical tube into 450 mL Millipore water with stirring. pH to 8.0
Read more

Laboratory Maintenance Checklist

-
Image
It is a known fact for anyone working in a laboratory be it the students, the researchers, scientists or lab assistants, it is necessary to follow safety rules and guidelines for their own safety. All laboratories have their lab safety symbols and lab safety equipments like goggles, mask, dress etc. At the same time, it is also very essential that a laboratory is well-maintained and taken care of so as to prevent any kind of dangers involved while working in a lab. As per rule, there should be routine maintenance and safety checks in a laboratory. Infact, it should be the duty of every person working in a laboratory to do a quick safety check after the completion of each experiment and before the starting of a new one. Or a responsible personnel should be assigned the duty to clean the laboratory and perform simple maintenance for lab care and safety . Laboratory Maintenance Presenting below few points which should be considered for laboratory maintenance: Bench:
Read more