Freezedown Guidelines
互联网
Freezedown Guidelines
What to Save
For each 96-pack, each lab will have yeast representatives from all four strains: BY4741 (or BY4730), BY4742 (or BY4739), BY4743, and the homozygous diploid. It is recognized that these four collections will each have gaps, due to (a) some genes being essential, (b) failure to obtain transformants that passed all 5 PCR confirmation tests.
Central Storage:
We will be storing 4 sets of A isolates in 2-ml tubes.
For essential genes only we will store the B isolates in addition to the A isolates.
Two sets go to Research Genetics, one each to Stanford and EUROSCARF.
Duplicates of 'A' strains, 'B' isolates , etc. may be saved according to each lab's preferences
Send strains off when you're done with the 96-pack and have entered all the data for those strains in the database.
Storage in 2-ml tubes:
-
Patch strain out on a YPD-G418 plate.
-
Color-code the caps of the Sarstedt tubes with a VWR pen or equivalent --
BY4741 or BY4730 -The MAT a haploid-RED
BY4742 or BY4739- The MAT alpha haploid-BLUE
BY4743 - The heterozygous diploid- BLACK
The homozygous diploid-GREEN -
Pick yeast up with a toothpick or implement of your choice and transfer to 1.0 or 1.5 ml YPD/15% glycerol in a Sarstedt tube. Transfer enough yeast into the tube to get an OD600 of about 5 (estimate by eye).
-
Freeze at -70° C before cells the settle.
-
One 100 place box should be used per background per 96-pack, regardless of the fact that the boxes will have gaps.
-
Put the B isolates of the essential genes either in a separate box or at the "end" of the homozygous diploid box.
-
Send on dry ice.
-
Notify the people you're sending them to by e-mail
Tubes: 2-ml tubes with writing area that fit 100/2" box (Sarstedt #72.694.006)
Labels : You will receive printed out labels for 6 sets of tubes from Stanford . If you start freezing before these are available, label each tube with the record number, strain name, batch and 96-pack co-ordinate on the side.
Boxes : National Lab Net, 908 549 2100, cat# R8300
