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--- sample_collection_and_dna_extraction [2012/02/22 12:03]
anniearchambault
+++ sample_collection_and_dna_extraction [2012/02/22 14:28]
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 ==Environmental or fungal samples==
-For environmental samples, such as soil core or rhizosphere, a variety of methods exist for preserving DNA integrity between field sampling and laboratory DNA extraction. When compared, the non-toxic sucrose lysis buffer (SLB) surpassed the guanidine isothiocyanate (GIT) buffer for preserving DNA from sediment samples stored at room temperature for a week Mitchell and Takacs-Vesbach 2008 ((Mitchell, K.R., Takacs-Vesbach, C.D., 2008. A comparison of methods for total community DNA preservation and extraction from various thermal environments. Journal of Industrial Microbiology & Biotechnology 35, [[http://www.springerlink.com/content/w638035028100452/|1139–1147. 10.1007/s10295-008-0393-y]])). Other sample storage buffers available are a CTAB buffer in saturated NaCl, 200 mM sodium ascorbate on samples previously homogenized in a sorbitol wash buffer ; or a TNES buffer Nagy 2010((Nagy, Z.T., 2010. A hands-on overview of tissue preservation methods for molecular genetic analyses. Organisms Diversity & Evolution 10, 91–105[[http://www.springerlink.com/content/c81h415005u27534/fulltext.html|10.1007/s13127-010-0012-4]])).
+For environmental samples, such as soil core or rhizosphere, a variety of methods exist for preserving DNA integrity between field sampling and laboratory DNA extraction. When compared, the non-toxic sucrose lysis buffer (SLB) surpassed the guanidine isothiocyanate (GIT) buffer for preserving DNA from sediment samples stored at room temperature for a week Mitchell and Takacs-Vesbach 2008 ((Mitchell, K.R., Takacs-Vesbach, C.D., 2008. A comparison of methods for total community DNA preservation and extraction from various thermal environments. Journal of Industrial Microbiology & Biotechnology 35, [[http://www.springerlink.com/content/w638035028100452/|1139–1147. 10.1007/s10295-008-0393-y]])). Other sample storage buffers available are a CTAB buffer in saturated NaCl, 200 mM sodium ascorbate on samples previously homogenized in a sorbitol wash buffer ; or a TNES buffer (Nagy 2010((Nagy, Z.T., 2010. A hands-on overview of tissue preservation methods for molecular genetic analyses. Organisms Diversity & Evolution 10, 91–105[[http://www.springerlink.com/content/c81h415005u27534/fulltext.html|10.1007/s13127-010-0012-4]])) ).
-Many QCBS researchers routinely analyze genetic diversity of fungi and of microorganisms from environmental samples. When based in the arctic, permafrost core are simply maintained frozen until DNA extraction can take place ([[http://qcbs.ca/members/main-researchers/?profile=68|Lyle Whyte]]). In warmer areas, soil samples are commonly sieved on site, kept in plastic bag for the day, then frozen ([[http://qcbs.ca/members/main-researchers/?profile=72|Yolande Dalpé]], [[http://qcbs.ca/members/main-researchers/?profile=42|Mohamed Hijri]], [[http://qcbs.ca/members/main-researchers/?profile=60|Marc St-Arnaud]]).
+Many QCBS researchers routinely analyze genetic diversity of fungi and of microorganisms from environmental samples. When based in the arctic, permafrost core are simply maintained frozen until DNA extraction can take place ([[http://qcbs.ca/members/main-researchers/?profile=68|Lyle Whyte]]). In warmer areas, soil samples are commonly sieved on site, kept in plastic bag on ice packs in a cooler for the day, then frozen ([[http://qcbs.ca/members/main-researchers/?profile=72|Yolande Dalpé]], [[http://qcbs.ca/members/main-researchers/?profile=42|Mohamed Hijri]], [[http://qcbs.ca/members/main-researchers/?profile=60|Marc St-Arnaud]]). Samples can also be instantly frozen in ethanol poured over on dry ice, in a cooler, or preserved in a commercial  [[http://www.mobio.com/soil-rna-isolation/lifeguard-soil-preservation-solution--100ml.html|LifeGuard™ Soil Preservation Solution]] that will enable separation of soil from rhizosphere.