| 查看: 1833 | 回复: 5 | |||
sihuanchen新虫 (小有名气)
|
[交流]
【求助/交流】5-氟乳清酸培养基 已有2人参与
|
| 5-氟乳清酸培养基配制，是先加5-氟乳清酸再灭菌还是灭菌后再加？ |
回复此楼» 本帖已获得的红花（最新10朵）
lyyjackey» 猜你喜欢
不合理蛙科研实验中的趣事：实验器材的 “乌龙”
已经有0人回复
-
不合理蛙科研实验中的趣事：和实验材料的 “斗智斗勇”
已经有0人回复
-
化学工程及工业化学论文润色/翻译怎么收费?
已经有226人回复
-
蛋白质检测：精准分析，解锁生物分子的密码
已经有0人回复
-
不合理蛙科研实验之小鼠实验：严谨设计，解析生命机制的重要载体
已经有0人回复
-
不合理蛙科研实验之重金属检测：精准筛查，守护健康与环境的防线
已经有0人回复
-
不合理蛙科研实验之“蛙测重金属我背锅三千”
已经有0人回复
-
不合理蛙科研实验之“鼠逃三次我发三篇SCI”
已经有0人回复
-
纳米粒度分析
已经有3人回复
-
林科院林化所招收材料与化工专业硕士，坐标南京
已经有0人回复
» 本主题相关价值贴推荐，对您同样有帮助:
- 11月需回复帖汇总 欢迎广大热心虫子来应助交流 大量金币等你拿
已经有3人回复
- 【求助/交流】酿酒酵母URA3基因敲除的问题
已经有30人回复
- 【求助/交流】5-氟乳清酸的配制和灭菌
已经有3人回复
- 【求助】二氟二苯甲酮的磺化
已经有0人回复
lingn
木虫 (正式写手)
- 应助: 0 (幼儿园)
- 金币: 1785.8
- 散金: 1
- 帖子: 745
- 在线: 44.7小时
- 虫号: 1008653
- 注册: 2010-04-29
- 专业: 细胞增殖、生长与分化
2楼2010-05-16 20:06:41
Atalpha
木虫 (著名写手)
- 应助: 1 (幼儿园)
- 金币: 3261.9
- 红花: 4
- 帖子: 1684
- 在线: 131.4小时
- 虫号: 925795
- 注册: 2009-12-11
- 性别: GG
- 专业: 植物学
★ ★ ★ ★
sihuanchen(金币+1):谢谢参与
amisking(金币+3):good！ 2010-05-16 21:52:28
sihuanchen(金币+20): 2010-08-24 17:08:43
sihuanchen(金币+1):谢谢参与
amisking(金币+3):good！ 2010-05-16 21:52:28
sihuanchen(金币+20): 2010-08-24 17:08:43
|
琼脂培养基先灭菌冷却到50-60度时再加入用0.2 um滤膜过滤除菌的5-氟乳清酸，提醒一下，5-氟乳清酸再水里，甲、乙醇都很难溶解，在水里溶解要加热到50度左右搅拌一个小时左右才充分溶解，溶解后用0.2 um滤膜过滤除菌。我把我用的protocol给您贴出来吧！ Good luck with your experiments! 5-FOA (5-Fluoroorotic Acid) Medium and Agar Plate Recipe Following recipe makes 1 liter 5-FOA plates: Step-1: Autoclave 20 grams of agar in 750 ml double distilled water. Step-2: Add followings into a glass beaker, and bring to 250 ml with double distilled water. Ammonium Sulfate 5.0 g Yeast nitrogen base without amino acid 1.7 g Dextrose 20 g Necessary nutritional supplements (see note 3) X g 5-FOA 1.0 g Warm up 5-FOA mixture in Step-2 at 55-65°C on a heat plate, stir until it dissolves ~30 min-1 h. The solubility of 5-FOA is very low in water; therefore, it is necessary to warm up the solution. Some manufacturers, i.e., US Biological, claim that 5-FOA is resistant to autoclaving. I did not try to autoclave it myself, but you can try it. After 5-FOA containing mixture in Step-2 is completely dissolved, sterilize it by filter. Place both melted agar and 5-FOA containing solution in a water bath (50-60°C) for one hour. Finally, add 5-FOA solution onto melted agar and pour your plates. Plates may be dried at RT for 48 hours and may be kept at 4°C for long term storage. Notes: 1.Do not forget to add Uracil into the plates at 20 mg/L of final concentration (Do not add too much Uracil!). 2.Never adjust the pH of 5-FOA containing medium. Only protonated form of 5-FOA is permeable. pH of your plates will be very acidic after adding 5-FOA. 3.Do not add more nutritional supplements than it is necessary; this reduces efficacy of 5-FOA plates. For example, you only need to add His, Leu, Lys and Ura supplements for strain BY4742. These supplements may be added directly from 100x stock solution. 4.5-FOA is very expensive reagent. I found that US Biological has the cheapest price for 5-FOA. 5.5-FOA is highly mutagenic to Candida albicans. This may also be the case for budding yeast. I suggest you do not leave yeast colonies on 5-FOA plates for longer than it is necessary. Wellington and Rustchenko, Yeast, 2005, Volume 22, Issue 1, Pages 57-70). Other sources of 5-FOA is Zymo Research and Sigma. The US Biological sells the cheapest 5-FOA reagent. I do not know if purity of 5-FOA is different from each company. 5-FOA from US Biological worked great for me. Do not waste your money! Good luck with your experiments! |
3楼2010-05-16 20:21:30
yatou996512
新虫 (小有名气)
- 应助: 1 (幼儿园)
- 金币: 96.7
- 散金: 28
- 帖子: 180
- 在线: 79.1小时
- 虫号: 1019671
- 注册: 2010-05-16
- 专业: 基础兽医学
4楼2011-10-10 16:36:06
5楼2013-05-21 20:07:35
loveqiuqiu
铜虫 (小有名气)
- 应助: 2 (幼儿园)
- 金币: 135.3
- 红花: 1
- 帖子: 69
- 在线: 10.4小时
- 虫号: 3592338
- 注册: 2014-12-13
- 性别: MM
- 专业: 微生物遗传学
6楼2015-01-17 21:13:21
