DAVID,
IN MAY, VIC DEFILIPPIS ASKED IF ANYONE HAD HAD ANY PROBLEMS REUSING
CENTERI-SEP COLUMNS. I RAN INTO A SIMILIAR PROBLEM BUT JUST IN A FEW
COLUMNS. I WOULD STORE THE USED COLUMNS IN WATER AND HAVE A LAB HELPER
SPIN WATER THROUGH THEM 3X'S AND LET DRY FOR A COUPLE OF DAYS BEFORE
LOADING AGAIN. WE FINALLY JUST STARTING KEEPING TRACK OF HOW OFTER THEY
WERE USED AND THREW THEM OUT AFTER THE 3RD REUSE. FORTUNATELY, I HAD TWO
OTHER CORE OPPERATORS THAT DIDN'T REUSE THE COLUMNS AND WOULD SAVE ME THEIR
OLD COLUMNS FOR ME.
I DID STOP USING CENTRI-SEP COMPLETELY DUE TO COST WHEN I FOUND BIOEDGE'S
96 WELL BLOCKS. THE COST PER WELL ON WITH THE BLOCKS IS ABOUT $0.70. IF A
WHOLE BLOCK IS NOT NECESSARY, I CUT THE SEAL BETWEEN COLUMNS AND SPIN THE
BLOCK AS USUAL. I PUT THE BLOCK BACK AT 4°C UNTIL I NEED IT. I DON'T DO
ANYTHING WHEN I GO TO USE THE STORED BLOCK, JUST TREAT IT LIKE AN UNUSED
BLOCK. I'VE HAD NO PROBLEM WITH STRETCHING OUT THE BLOCKS, EVEN WHEN SOME
HAVE BEEN STORED UP TO TWO WEEKS.
WHAT I MISS ABOUT NEW CENTRI-SEPS, IS THAT I FEEL THE RECOVERY OF
SEQUENCING PRODUCT IS SO MUCH BETTER. I CAN EASILY TELL THE DIFFERENCE ON
THE GEL IMAGE. IF I HAVE A COUPLE OF SAMPLES THAT HAVE TO "CATCH UP" WITH
THE OTHER SAMPLES FOR A DAY'S RUN, I WILL USUALLY USE CENTRI-SEP COLUMNS
AND THOSE SAMPLE WILL ALWAYS COME BACK WITH BIGGER PELLETS AND BRIGHTER
BANDS. THE SEQUENCE QUALITY USING THE BIOEDGE BLOCK DOESN'T SEEM TO SUFFER
THOUGH.
RUNNING 96/DAY THE CENTRI-SEP ARE JUST TOO TIME CONSUMING AND EXPENSIVE @
>2.00/COLUMN. THE PRINSTON SEPARATIONS REP DID COME BY WITH THEIR VERSION
OF THE 96WELL BLOCK BUT IT WAS STILL ABOUT $0.15 MORE PER WELL THAN BIOEDGE
AND LESS EASY TO USE. I DID TEST CUTTTING THE SEAL ON THOSE TOO AND HAD NO
PROBLEM STORING AND USING THEM LATER.
SHIRLEY HALL
UT SOUTHWESTERN MEDICAL CENTER
DEPT OF MOLECULAR BIOLOGY
