20.109-Protocols/Thawing J1

Similar to that described on atcc.org for product SCRC-1010, with some changes. Briefly,


 * 1) Coat the culture flask to be used with pre-warmed gelatin for at least 10 min.
 * 2) *Use 5 mL for a T75, or 1 mL for one well of a 6-well plate.
 * 3) Frozen J1 are stored in the Niles lab liquid nitrogen vessel. Bring an aliquot to lab, and thaw it in the 37 &deg;C water bath.
 * 4) *Gently shake the vial, while submerging only the body - keep the cap above the surface of the water to prevent contamination.
 * 5) *Limit the immersion time to the minimum needed (~2 min).
 * 6) Pipet the 1 mL of cells into a conical tube containing 8 mL of pre-warmed culture medium. Let the cells fall drop by drop.
 * 7) To wash any remaining cells out of the vial, use another 1 mL of medium.
 * 8) Spin the cells down for 5 min at 500 g.
 * 9) *Our centrifuge does not work well at lower rcf.
 * 10) Resuspend the cells in an appropriate volume.
 * 11) *For a T75, culture in 15 mL total. For one well of a 6-well, use 6 mL.
 * 12) If you use a T75...
 * 13) *Change the medium every day.
 * 14) *A 5M aliquot will probably need about 3-4 d of growth before splitting.
 * 15) If you use a 6-well...
 * 16) *Change the medium after the first day.
 * 17) *A 5M aliquot will probably need about 1-2 d of growth before splitting to a single T75.
 * 18) *After 1-3 days, the cells should be propagating normally and ready for a typical split.