Success with DNA

On Monday, we discussed our IJSEM paper. Individually, we will attempt to describe a new species from our 26 stains within the family Geodermatophilaceae. Then our desiccation experiment from February 9 was ready to be spotted onto media. 500µl of liquid media was added to each well on the desiccation tray. After thirty minutes, each well was gently mixed with a pipette for 30 seconds, then 10µl were spotted onto media. Then the PCR was set up with GEO primers for our DNA that was extracted from the dirt samples last week. PCR was set up a little different this week, by adding the Taq after the samples were heated at 98ºC for ten minutes. Also new stock plates were restreaked including our GB-20 strain that had positive PCR results. Then we recorded our 9 kGy results into the spread sheet. We found that five of our strains were able to tolerate this high irradiation.
The pictures show some of our stains that are tolerant of 9kGy.

Strain 67 and 72 both seem almost non-affected by the irradiation.
Strain 68 grew but showed less growth after the irradiation.
Tuesday, Eugene ran a gel with our PCR results from Monday and number of other dirt and bacteria samples. The results from out dirt samples showed a strong presence for bacteria from the family Geodermatophilaceae with bright bands around 565 bps.
Wednesday, we spotted of strains on 17 new carbon sources which included:
Fructose FR
Cellobiose CB
Pyruvate PY
Trehalose TR
Sorbitol SB
Myo-inositol MI
Maltose MT
Citrate CT
Acetate AC
Succinate SC
Mannose MS
Alanine AL
Arginine AG
Asparagine AS
Histidine HS
Lysine LY
Threonine TH
We also purified eight of Dr. Rainey’s PCR samples following the PCR purification protocol from Mo Bio. Six out of eight of these samples were also a success on a gel.