1 / 21

Molecular Methods in Bioremediation

Molecular Methods in Bioremediation Polymerase Chain Reaction (PCR) PCR Machine Gel Electrophoresis Primer Primer sequence Annealing Size Reference temperatures ( C) 16S rDNA universal-F 5 ′ -CCGTCGACAGAGTTTGATCCTGGCTCAG- 3 ′ 55 1500 1 16S rDNA universal-R

niveditha
Télécharger la présentation

Molecular Methods in Bioremediation

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Molecular Methods in Bioremediation

  2. Polymerase Chain Reaction (PCR)

  3. PCR Machine

  4. Gel Electrophoresis

  5. Primer Primer sequence Annealing Size Reference temperatures (C) 16S rDNA universal-F 5′-CCGTCGACAGAGTTTGATCCTGGCTCAG-3′ 55 1500 1 16S rDNA universal-R 5′-CGGGATCCACCTTGTTACGACTTCACCC-3′ 55 1 Bacillus sp. -R 5′-CCCAGTTTCCAATGACC-3′ 55 640 1 Pseudomonas sp. -R 5′-CCTTCCTCCCAACTT-3′ 50 440 2 Catabolic gene alkM-F 5′-CGGCTACTCCGATGATCGG-3′ 62 870 3 Table 1. Oligonucleotide primers used to identified hydrocarbon degrading bacteria alkM-R 5′-GATCCGAGTGCCGCTGAAG-3′ 62 3 ndoM-F 5′-CACTCATGATAGCCTGATTCC-3′ 62 642 3 ndoM-R 5′-CACAACACACCCATGCCGCTG-3′ 62 3 todM-F 5′-GGGCTTACGACACCGCCG-3′ 62 560 3 todM-R 5′-GCGCTCCACGCTACCCAG-3′ 62 3 xylM-F 5′-CTGCGTGTACTGGACATGAG-3′ 62 834 3 xylM-R 5′-CGGTGGTCCAGGTCACCG-3′ 62 3 1—Arellano and Olmos, 2002; 2—Personal communication (Sandra Nierzwicki Bauer); Olmos, 2003; 3—The present work, primers modified from Whyte et al. (1996). Oligos for Hydrocarbon Degraders

  6. Detection by PCR Amplification Lane 1: Molecular Weight Marker Lanes 2-8: PCR products from DNA extracted for soil suspensions using Pseudomonas primers Lanes 9-15: Bacillus primers

  7. DNA Hybridization

  8. DNA Sequencing

  9. TRFLP

  10. Real-time TRFLP Yu, et al. AEM 71:1433-1444

  11. DGGE

  12. Genome Sequencing

  13. Microarrays

  14. TaqMan Real-time PCR Images by Dan Pierce

  15. Image by Dan Pierce http://www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Pierce/realtimepcr.htm

  16. Other Techniques • Stable Isotope Probing (SIP) • Fluorescent In Situ Hybridization (FISH) • mRNA Analysis

More Related