Soil Sample 5508 / Culture 10518 / Isolate 10519
Isolate 10519
MGKA3 University of Connecticut
Published
ESKAPE Test
Published
Recorded Friday, December 6, 2019
Tested against ESKAPE relative Antibiotic activity Length of incubation Media used
Bacillus subtilis Yes 24.0 hr 10% Tryptic Soy Agar (TSA)
Enterococcus raffinosus No
Staphylococcus epidermidis Yes 24.0 hr 10% Tryptic Soy Agar (TSA)
Escherichia coli No
Acinetobacter baylyi No
Pseudomonas putida No
Enterbacter aerogenes No
Mycobacterium smegmatis Yes 24.0 hr 10% Tryptic Soy Agar (TSA)
16S rRNA Test
Published
Recorded Friday, December 6, 2019
Genus
Pseudomonas
FASTA sequence
MGKA3_r-519R.seq DOWNLOAD FILE
MGKA3_f-27F.seq DOWNLOAD FILE
Distinguishing Characteristics
Antibiotic Activity of Extract
Published
Recorded Friday, December 6, 2019
Solvent used
Methanol
Experiment Description
Metabolite extraction  -obtain Swab plates of isolates  -using a sterile spatula cut the agar into 1mm cubes -place cut up agar into screw-top jar and label with respective isolate names  -place screw top jar containing agar into cooler with a dry ice and ethanol bath  -allow a deep freeze of agar for at least 20 minutes  -remove screw top jar from dry ice and ethanol bath under a hood add 15ml of ethyl acetate and 10ml of  distilled water to jar.  -allow mixture to incubate on a shaker for 24 hours -obtain two scintillation vials for each isolate -label one of the  vials for each isolate extract and measure and record the mass of the empty vial -remove mixture in screw top jars from incubation  -using a sterile transfer pipette transfer all liquid in the screw top jar to the unlabeled scintillation vial  -allow liquid to settle in scintillation jar  -using another sterile pipette transfer the top layer of settled solution into the labeled scintillation vial  -let vial sit opened in hood to dry down over a 3-4 day period  -After solution has dried down  measure and record the mass of the scintillation vial -Add 1ml of methanol to scintillation vial and record the concentration  -use an ultrasonic cleaner in order to homogenize mixture -perform dilutions as necessary to get concentrations of 10mg/ml, 25mg/ml and 50mg/ml Chia seeds -Obtain and label water agar plates for each of the isolate extracts at concentrations of 10mg/ml, 25mg/ml and 50mg/ml to be tested positive control using tetracycline at a concentration of 5mg/ml. And negative control of methanol -Draw circles on the bottom of the plates for each treatment -using forceps place 5 chia seeds in each circle -Pipette 10 microliters of each treatment into their respective labeled circles. -Repeat above step three times for each extract allowing it to dry in between -Incubate water agar plates at room temperature with adequate sunlight
Tested against ESKAPE relative Antibiotic activity Length of incubation Media used
Bacillus subtilis No
Enterococcus raffinosus No
Staphylococcus epidermidis Yes 24.0 hr LB
Escherichia coli No
Acinetobacter baylyi No
Pseudomonas putida No
Enterbacter aerogenes No
Mycobacterium smegmatis Yes 24.0 hr LB
Eukaryotic Inhibition
Published
Recorded Friday, December 6, 2019
Experiment description
Metabolite extraction  -obtain Swab plates of isolates  -using a sterile spatula cut the agar into 1mm cubes -place cut up agar into screw-top jar and label with respective isolate names  -place screw top jar containing agar into cooler with a dry ice and ethanol bath  -allow a deep freeze of agar for at least 20 minutes  -remove screw top jar from dry ice and ethanol bath under a hood add 15ml of ethyl acetate and 10ml of  distilled water to jar.  -allow mixture to incubate on a shaker for 24 hours -obtain two scintillation vials for each isolate -label one of the  vials for each isolate extract and measure and record the mass of the empty vial -remove mixture in screw top jars from incubation  -using a sterile transfer pipette transfer all liquid in the screw top jar to the unlabeled scintillation vial  -allow liquid to settle in scintillation jar  -using another sterile pipette transfer the top layer of settled solution into the labeled scintillation vial  -let vial sit opened in hood to dry down over a 3-4 day period  -After solution has dried down  measure and record the mass of the scintillation vial -Add 1ml of methanol to scintillation vial and record the concentration  -use an ultrasonic cleaner in order to homogenize mixture -perform dilutions as necessary to get concentrations of 10mg/ml, 25mg/ml and 50mg/ml Chia seeds -Obtain and label water agar plates for each of the isolate extracts at concentrations of 10mg/ml, 25mg/ml and 50mg/ml to be tested positive control using tetracycline at a concentration of 5mg/ml. And negative control of methanol -Draw circles on the bottom of the plates for each treatment -using forceps place 5 chia seeds in each circle -Pipette 10 microliters of each treatment into their respective labeled circles. -Repeat above step three times for each extract allowing it to dry in between -Incubate water agar plates at room temperature with adequate sunlight
Eukaryotic organism Eukaryotic inhibition
Chia seeds No
Associated Entries

Soil Sample 5508
Collected Thursday August 29, 2019

Published

Culture Conditions 10520
Recorded Friday December 6, 2019

Published

Isolate 10521
Recorded Friday December 6, 2019

Published

Culture Conditions 10518
Recorded Friday December 6, 2019

Published

Isolate 10519
Recorded Friday December 6, 2019

Published