Molecular methods in bioremediation
Download
1 / 21

Molecular Methods in Bioremediation - PowerPoint PPT Presentation


  • 597 Views
  • Updated On :

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

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'Molecular Methods in Bioremediation' - niveditha


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.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript



Gel electrophoresis l.jpg
Gel Electrophoresis


Oligos for hydrocarbon degraders l.jpg

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


Detection by pcr amplification l.jpg
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



Dna sequencing l.jpg
DNA Sequencing



Real time trflp l.jpg
Real-time TRFLP

Yu, et al. AEM 71:1433-1444



Genome sequencing l.jpg
Genome Sequencing



Taqman real time pcr l.jpg
TaqMan Real-time PCR

Images by Dan Pierce


Slide19 l.jpg

Image by Dan Pierce

http://www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Pierce/realtimepcr.htm


Other techniques l.jpg
Other Techniques

  • Stable Isotope Probing (SIP)

  • Fluorescent In Situ Hybridization (FISH)

  • mRNA Analysis


ad