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How can I tell if there is contamination?

How can I tell if there is contamination?

Biology: The Unity and Diversity of Life (MindTap Course List)
Biology: The Unity and Diversity of Life (MindTap Course List)
15th Edition
ISBN:9781337408332
Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Chapter46: Ecosystems
Section: Chapter Questions
Problem 3CT: Nitrogen-fixing bacteria live throughout the ocean, from its sunlit upper waters to 200 meters (650...
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How can I tell if there is contamination?
Results to turn in Identification of Enterococcus Species from Beach SandSamples Your goal is to use the PCR-RFLP procedure with gene cloning to determine if ENT species are present and if so, are their sub-species within the samples. You are also interested in determining whether there are differential spatial profiles of ENT bacteria at the sampling sites. In particular, you want to determine if the proximity to the storm drain outfall is a source of sand bacterial contamination. -360CG forward primer reverse primer Kas I (GGCGCC) RFLP -360C -360G GGCGCC GGGGCC 214 bp 276 bp 93 bp 490 bp 93 bp м 1 3 4. 5 6 8 9 10 11 12 C2 500 bp 400 bp +490 bp 300 bp -276 bp 214bp 200 bp -93 bp C, Fig. 3. PCR-RFLP for genotyping -360C/G in the gene encoding deoxycytidine kinase. In the upper part of the figure, the amplified fragment and the restriction fragments for the two alleles produced by treatment with Kasl are shown. Note that the amplified fragment contains a non-polymorphic Kasl recognition site. Fragment sizes are 214, 276 and 93 bp for the Kasl- positive allele and 490 and 93 bp for the other allele. In the lower part of the figure, the restriction fragments are shown in gel view format. There is one heterozygote (lane 4); all others are homozygotes. Cl and C2 are internal calibration markers. Adapted with permission from Szantai et al., (2006).
Transcribed Image Text:Results to turn in Identification of Enterococcus Species from Beach SandSamples Your goal is to use the PCR-RFLP procedure with gene cloning to determine if ENT species are present and if so, are their sub-species within the samples. You are also interested in determining whether there are differential spatial profiles of ENT bacteria at the sampling sites. In particular, you want to determine if the proximity to the storm drain outfall is a source of sand bacterial contamination. -360CG forward primer reverse primer Kas I (GGCGCC) RFLP -360C -360G GGCGCC GGGGCC 214 bp 276 bp 93 bp 490 bp 93 bp м 1 3 4. 5 6 8 9 10 11 12 C2 500 bp 400 bp +490 bp 300 bp -276 bp 214bp 200 bp -93 bp C, Fig. 3. PCR-RFLP for genotyping -360C/G in the gene encoding deoxycytidine kinase. In the upper part of the figure, the amplified fragment and the restriction fragments for the two alleles produced by treatment with Kasl are shown. Note that the amplified fragment contains a non-polymorphic Kasl recognition site. Fragment sizes are 214, 276 and 93 bp for the Kasl- positive allele and 490 and 93 bp for the other allele. In the lower part of the figure, the restriction fragments are shown in gel view format. There is one heterozygote (lane 4); all others are homozygotes. Cl and C2 are internal calibration markers. Adapted with permission from Szantai et al., (2006).
O Untitl. E Un 8 μΙ f 10 µl 10 µl 10 μι Sample 2 Sample 3 20ul 24 µl of Mlyl cut 24 µl of 24 µl of DNA Sample 1 Sample 4 PCR + 2 µl PCR + 2 µl PCR + 2 µl PCR (H2O Sample 1 Sample 2 Sample 3 Mlyl cut Mlyl cut ladder loading loading loading control) + 4 µl PCR PCR PCR product product product loading Are Sample A) Is there contamination in any of the sand samples (S1, S2, S3) B) If contaminated from analyzing restriction digest band patterns, what is the contaminating species?
Transcribed Image Text:O Untitl. E Un 8 μΙ f 10 µl 10 µl 10 μι Sample 2 Sample 3 20ul 24 µl of Mlyl cut 24 µl of 24 µl of DNA Sample 1 Sample 4 PCR + 2 µl PCR + 2 µl PCR + 2 µl PCR (H2O Sample 1 Sample 2 Sample 3 Mlyl cut Mlyl cut ladder loading loading loading control) + 4 µl PCR PCR PCR product product product loading Are Sample A) Is there contamination in any of the sand samples (S1, S2, S3) B) If contaminated from analyzing restriction digest band patterns, what is the contaminating species?
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