黄色小说视频-日本少妇高潮抽搐-国内自拍av-天堂中文资源在线-蜜桃视频在线入口www-91久久综合-亚洲视频在线一区-日日夜夜拍-国产精品资源在线观看-欧美激情国产精品免费-人妻少妇久久中文字幕-成人免费无码大片a毛片-香蕉污视频在线观看-日本电影成人-国内精品久久99人妻无码

Yes

1. Harvest cells from a culture which is at or near peak density by centrifugation at ~800 x g for 10 min.
2. Adjust concentration of cells to 2 x 10⁷ - 2 x 10⁸ mL in fresh growth medium.  If the concentration is too low, centrifuge at ~800 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired cell concentration.
3. While cells are centrifuging, prepare a 10% (v/v) solution of sterile methanol in fresh growth medium. 
4. Mix the cell suspension and the methanol solution in equal portions. The final concentration will be 10⁷ – 10⁸ cells/mL and 5% (v/v) of methanol. The time from the mixing of the cell preparation and methanol stock solution before the freezing process is begun should be no less than 5 min and no more than 15 min.
5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.
6. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion.  At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)
7. Store in either the vapor or liquid phase of a nitrogen refrigerator.
8. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after it is thawed.
9. Remove the vial from the water bath immediately after thawing.  Aseptically transfer the contents of the ampule into 5 mL of fresh ATCC medium 2861. 
10. Incubate the tube on a 15^° horizontal slant with the cap screwed on loosely (loosened one half turn) at 42°C under constant light.
11. Maintain as described above.  

Ohta N, et al. Structure and organization of the mitochondrial genome of the unicellular red alga Cyanidioschyzon merolae deduced from the complete nucleotide sequence. Nucleic Acids Res. 26(22): 5190-5198, 1998. PubMed: 9801318

Ohta N, et al. Complete sequence and analysis of the plastid genome of the unicellular red alga Cyanidioschyzon merolae. DNA Res. 10(2): 67-77, 2003. PubMed: 12755171

Matsuzaki M, et al. Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D. Nature 428(6983): 653-657, 2004. PubMed: 15071595

Nozaki H, et al. A 100%-complete sequence reveals unusually simple genomic features in the hot-spring red alga Cyanidioschyzon merolae. BMC Biol. 5: 28, 2007. PubMed: 17623057