Debra Medina Autoclave Sterilizer

Thermal Cycler

GENOMIC CHARACTRIZATION OF AEROMONAS HYDROPHILA ISOLATES USING RAPD-PCR TECHNIQUE by Mitali Dhiman and S. S. Mishra Central Inland Fisheries Resear

 

Aeromonas hydrophila are  involved in various disease problems in humans and aquatic  animals and are known to be  phenotypically, serologically and genetically quite diverse. Development and use of a sensitive and specific  diagnostic test is warranted for detection and characterization of this pathogen.In the present study,  Fish  and water samples from river Hooghly were streaked onto Aeromonas selective growth medium (Rimler-shotts Medium, HiMedia, Mumbai). After incubation at 30oC for 24 hrs  plates were observed for characteristic colony development. The bacteria were then identified using a battery of  standard cultural and biochemical tests (Buchanan and Gibbons., 1988). Bacterial DNA  was extracted following the method of  Sambrook et al., (1989). Primers specific for ‘aerolysin' gene (209 bp product, Ozbas et al., 2000) were used as the target gene for PCR amplification.  PCR mix of 50 µl containing 1.0U Taq polymerase, 5 μl 10X assay buffer, 0.1μm each of forward and reverse primers, 200 μm dNTPs and 1μl bacterial DNA sample was amplified using Thermal cycler (Gene Amp2400 PCR system, Applied Biosystem).  After 35 cycles of amplification (at 94oC 2 min, 56oC 2 min and 72o C 2 min), the products were electrophoresed on 2% agarose gel and visualized under Gel documentation system. The sequence of primers used was : Forward primer - 5'- CCA-AGG-GGT-CTG-TGG-CGA-CA-3', Reverse primer -5'- TTT-CAC-CGG-TAA-CAG-GAT-TG-3'.

 Selected A.hydrophila  isolates were used in RAPD-PCR analysis. First, a set of 20 oligo primers (Operon A & B set)  were screened  to see the suitability of primers in producing polymorphic bands. The amplification was carried out using Thermal cycler  using  the following programme :  one cycle of initial denaturation at 94oC x 5 min. followed by 45 cycles at 94oC for 1min., 36oC x1min. and 72oC x 2min. and final extension at 72oCx10 min.  PCR amplified products were analyzed on 1.5% agarose gel (Sambrook et al., 1989). Accordingly 2 primers (OPA5 and OPA 9), which produced good polymorphic bands were selected and used in RAPD-PCR analysis of  all A.hydrophila. The banding pattern and  RAPD profiles were analyzed  using the Gel documentation system (Geldoc 2000, BioRad) and  TFPGA software.

Results and Discussions

 Out of 32  isolates on RS medium screened, 20  were identified as  Aeromonas species. However, when these bacteria were  screened using  PCR for aerolysin gene, only 12 isolates and the standard MTCC strain  produced 209 bp specific DNA band indicating these to be A.hydrophila . No amplification  was seen in other  isolates and in negative controls. This indicated that PCR to be more specific and accurate for detection of A.hydrophila. Phylogenetic analysis of 12 A.hydrophila isolates was carried out using RAPD-PCR. Out of  20 primers screened  to test their suitability to produce  polymorphic  bands, 2 primers (OPA5 and OPA9) were selected and used for RAPD-PCR.  The banding pattern was estimated using the gel documentation system and the genetic similarity index was calculated as per  the formula of Nei and Li  using  TFPGA software.  Dendogram analysis of all isolates revealed high genetic variability among the isolates regardless of  their source of origin.  RAPD assays has been proved useful for identification and classification of a variety of microbial pathogens of humans and animals (Berg et al., 1994). However, RAPD profiles of A.hydrophila strains were scattered, demonstrating the genomic variety of motile aeromonads. There was no species specific fragments among profiles of A.hydrophila strains.

About the Author

Mitali Dhiman, Research Scholar. Dr.S.S.Mishra, Principal  Scientist,  Biotechnology Laboratory, Regional Centre,  Central Inland Fisheries  Research Institute, 24, Panna Lal Road, Allahabad - 211 002 (Uttar Pradesh) INDIA

Cycler $8.99 AS FAR AS anyone at her high school knows, Jill McTeague is an average smart girl trying to get her dream date to ask her to the prom. What no one knows, except for Jill’s mom and dad, is that for the four days Jill is out of school each month, she is not Jill at all. She is Jack, a genuine boy—complete with all the parts. Jack lives his four days per month in the solitude of Jill’s room. But his personality has been building since the cycling began. He is less and less content with his confinement and his cycles are becoming more frequent. Now Jill’s question about the prom isn’t who she'll go with, but who she'll be when the big night arrives. From the Hardcover edition.

Cycler's $18.48 No Synopsis Available

Artist's Concept of a Lunar Cycler $24.99 Stocktrek Images Artist's Concept of a Lunar Cycler - Photographic Print

Artist's Concept of a Lunar Cycler Approaching Earth $24.99 Stocktrek Images Artist's Concept of a Lunar Cycler Approaching Earth - Photographic Print

New Water Cycler Pac Aboveground Replacement Cartridge $54.99 New Water Cycler Pac / Replacement Cartridge for Aboveground PoolsThis is the Replacement Cartridge (Cycler Pac) for the New Water brand of aboveground swimming pool water mineral purifiers. Click here for Inground Cycler Pac.NOTES:This item is not compatible with Sparco, Prevail or Aqua Smarte systems.FedEx requires customer signature for delivery of this item.

New Water Cycler Pac Inground Replacement Cartridge $49.99 New Water Cycler Pac / Replacement Cartridge for Inground PoolsThis is the Replacement Cartridge (Cycler Pac) for the New Water brand of inground swimming pool water mineral purifiers. Click here for Aboveground Cycler Pac.NOTES:This item is not compatiblewith Sparco, Prevail or Aqua Smarte systems.FedEx requires customer signature for delivery of this item.

(Re)cycler $2.99 When eighteen-year-old Jill opts to move to New York with Ramie rather than take a road trip with her boyfriend Tommy, all of their relationships are shaken up as Jack, the boy she turns into for four days each month, finally has the chance to get to know himself.

New Water Cycler Pac Aboveground Replacement Cartridge - 6 Pack $199.99 New Water Cycler Pac Aboveground Replacement Cartridge - 6 Pack

New Water Cycler Pack Inground Replacement Cartridge - 6-Pack $249.99 New Water Cycler Pack Inground Replacement Cartridge - 6-Pack

Lone Cycler Race Scene Cigar Box Label, Cycling $19.99 Lone Cycler Race Scene Cigar Box Label, Cycling - Premium Poster

A Mars Cycler Travels by the Earth While Two Astronauts Watch from Afar $24.99 Stocktrek Images A Mars Cycler Travels by the Earth While Two Astronauts Watch from Afar - Photographic Print

A Manned Mars Lander/Return Vehicle Disembarks from a Mars Cycler $24.99 Stocktrek Images A Manned Mars Lander/Return Vehicle Disembarks from a Mars Cycler - Photographic Print

A Manned Mars Cycler Space Station Approaches the Planet Mars $24.99 Stocktrek Images A Manned Mars Cycler Space Station Approaches the Planet Mars - Photographic Print

Pool Frog Aboveground Cycler and Mineral Reservoir $179.99 Our Pool Frog Aboveground Mineral Package includes the Cycler, ReservoirWhy FROG systems are superior to Nature2® systems: Frog® mineral reservoirs use Ionic Silver and Nature2® uses Colloidal Silver. Ionic silver does NOT require a 2-step break-in, up to a 2-week period, to begin the mineral purification process. Frog begins to work the instant it is installed. No need to wait.You dont need to add chlorine or other sanitizers to activate the process. Nature2 needs the chlorine to break down the colloidal silver into ionic silver. Frog makes it easy to add the correct amount of chlorine to the pool with its Bac Pac design. A built in flow valve will always guarantee you will have the proper amount of minerals and sanitizer in the water to keep the water crystal clear, and swimmers, well protected. Once a season you can add a special algaecide called Bam utilizing the cycler feeder. This gives you a guarantee against algae for 90 days. Also, FROG is EPA registered as a sanitizer and Nature2 is EPA registered only as a disinfectant. Sanitizers kill, disinfectants might, but not like sanitizers do. Frog mineral pacs also incorporate a clarifier to help jump start your pool water for absolute clarity and brilliance. Frog ionic silver is coated on the outside of a calcium carbonate base and is delivered through via a time sequence mechanism. Frog never lets too much silver into the water. It is smart! Only the right amount of silver is ever in your pool to allow for optimum clarity and protection. Water is softer, no red eyes, no bleached suits, no green hair, Frog means you will work less and enjoy more!POOL FROG® ABOVE GROUND a complete system of Minerals and Chlorine, with a precise dial for achieving at least 0.5 ppm chlorine level. Includes a POOL FROG® Cycler 6100 and Mineral Reservoir Series 6112. Cycler is in-line with 1.5" threaded ports for pools up to 25,000 gallons. Mineral Reservoir lasts 6 months. New features include new easy to remove cap, no tools required, and new control dial for more precise feeding. More Fun With Pool FrogThe Above Ground POOL FROG® makes it even easier for you to enjoy your pool. The enhanced control dial allows for more precise control over the minerals and chlorine going into the pool. When its time to replace the minerals (every 6 months or once a season) or the chlorine POOL FROG®Bac Pac (every 2-3 weeks), the cap is a snap to remove. Simply grab the handles and twist it off. And these great features add to the cool look of the POOL FROG® Cycler.Put Frog Minerals To The TestYour pool will use less chlorine with far less work while still maintaining crystal clear water. Thats because POOL FROG® uses minerals that make the water feel softer and look sparkling clean.POOL FROG®s patented minerals destroy bacteria with a controlled release

Fisherbrand PCR Tubes, 0.2mL, Dome Caps, Natural, 1000/cs $46.37 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Dome Caps, Blue, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Dome Caps, Green, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Dome Caps, Red, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Dome Caps, Yellow, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Flat Caps, Natural, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Flat Caps, Blue, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Flat Caps, Green, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Flat Caps, Red, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Fisherbrand PCR Tubes, 0.2mL, Flat Caps, Yellow, 1000/cs $36.77 Features of the Fisherbrand PCR Tubes: With integral "snap shut" cap. Suitable for 0.2mL thermal cycler blocks. 0.25mL maximum volume when closed. Thin-wall design. Cleanroom produced. Virgin polypropylene.

Pool Frog Mineral Reservoir Pack for 5400 Inground Cycler $109.95 POOL FROG Mineral Reservoirs now make pool start up much easier with NEW START-UP WATER POLISHERS! This new feature added to the FROG® mineral technology gives the pool an extra boostof shine that helps start the season with the perfect pool. Now theres one more reason why POOL FROG® is the best option for above ground or in groundpools! This mineral reservoir is good for in-ground pools up to 40,000 gallons.This is the only mineral reservoir for the In-Ground Frog Cycler system.Looking for a Different Size or Model? Click here to view all related products.

Thermo Scientific ABgene 8-Strip Tubes, 0.2mL, Dome Caps, Natural, 80/cs $141.95 Features of the Thermo Scientific ABgene 8-Strip Tubes: Low-profile design reduces 'dead space' between cycler heated lid and sample, eliminating condensation on tube side, preventing reduction in PCR volume, and increasing reaction efficiency. Recommended for use with reaction volumes below 20?L. Suitable for 0.2mL thermal cycler blocks. 0.15mL maximum tube capacity when capped.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Assorted Colors, 1000/cs $66.19 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Natural, 1000/cs $48.21 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Blue, 1000/cs $57.65 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Green, 1000/cs $57.65 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Purple, 1000/cs $57.65 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Red, 1000/cs $57.65 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.

Thermo Scientific ABgene PCR Tubes, 0.2mL, Dome Caps, Yellow, 1000/cs $48.21 Features of the Thermo Scientific ABgene PCR Tubes: Suitable for PCR (0.2mL thermal cycler blocks). 0.25mL maximum tube capacity when closed. Thin-wall design, integral "snap shut" cap. Contamination-free virgin polypropylene.