 |
 |
 |
 |
 |
 |
||
 |
 |
 |
 |
 |
| 5 | 1/1 | 返回列表 |
| 查看: 1843 | 回復: 5 | |||
| 當前只顯示滿足指定條件的回帖，點擊這里查看本話題的所有回帖 | |||
sihuanchen新蟲 (小有名氣)
|
[交流]
【求助/交流】5-氟乳清酸培養(yǎng)基 已有2人參與
|
||
| 5-氟乳清酸培養(yǎng)基配制，是先加5-氟乳清酸再滅菌還是滅菌后再加？ |
回復此樓» 本帖已獲得的紅花（最新10朵）
lyyjackey» 猜你喜歡
不合理蛙科研實驗之小鼠實驗：嚴謹設計，解析生命機制的重要載體
已經(jīng)有0人回復
-
不合理蛙科研實驗之重金屬檢測：精準篩查，守護健康與環(huán)境的防線
已經(jīng)有0人回復
-
化學工程及工業(yè)化學論文潤色/翻譯怎么收費?
已經(jīng)有217人回復
-
不合理蛙科研實驗之“蛙測重金屬我背鍋三千”
已經(jīng)有0人回復
-
不合理蛙科研實驗之“鼠逃三次我發(fā)三篇SCI”
已經(jīng)有0人回復
-
納米粒度分析
已經(jīng)有3人回復
-
林科院林化所招收材料與化工專業(yè)碩士，坐標南京
已經(jīng)有0人回復
-
留學--博士招生
已經(jīng)有16人回復
-
化學工程，本211，求調劑，ab區(qū)不限
已經(jīng)有4人回復
-
邵陽學院食品與化學工程學院碩士調劑
已經(jīng)有0人回復
» 本主題相關價值貼推薦，對您同樣有幫助:
- 11月需回復帖匯總 歡迎廣大熱心蟲子來應助交流 大量金幣等你拿
已經(jīng)有3人回復
- 【求助/交流】釀酒酵母URA3基因敲除的問題
已經(jīng)有30人回復
- 【求助/交流】5-氟乳清酸的配制和滅菌
已經(jīng)有3人回復
- 【求助】二氟二苯甲酮的磺化
已經(jīng)有0人回復
yatou996512
新蟲 (小有名氣)
- 應助: 1 (幼兒園)
- 金幣: 96.7
- 散金: 28
- 帖子: 180
- 在線: 79.1小時
- 蟲號: 1019671
- 注冊: 2010-05-16
- 專業(yè): 基礎獸醫(yī)學
4樓2011-10-10 16:36:06
lingn
木蟲 (正式寫手)
- 應助: 0 (幼兒園)
- 金幣: 1785.8
- 散金: 1
- 帖子: 745
- 在線: 44.7小時
- 蟲號: 1008653
- 注冊: 2010-04-29
- 專業(yè): 細胞增殖、生長與分化
2樓2010-05-16 20:06:41
Atalpha
木蟲 (著名寫手)
- 應助: 1 (幼兒園)
- 金幣: 3261.9
- 紅花: 4
- 帖子: 1684
- 在線: 131.4小時
- 蟲號: 925795
- 注冊: 2009-12-11
- 性別: GG
- 專業(yè): 植物學
★ ★ ★ ★
sihuanchen(金幣+1):謝謝參與
amisking(金幣+3):good！ 2010-05-16 21:52:28
sihuanchen(金幣+20): 2010-08-24 17:08:43
sihuanchen(金幣+1):謝謝參與
amisking(金幣+3):good！ 2010-05-16 21:52:28
sihuanchen(金幣+20): 2010-08-24 17:08:43
|
瓊脂培養(yǎng)基先滅菌冷卻到50-60度時再加入用0.2 um濾膜過濾除菌的5-氟乳清酸，提醒一下，5-氟乳清酸再水里，甲、乙醇都很難溶解，在水里溶解要加熱到50度左右攪拌一個小時左右才充分溶解，溶解后用0.2 um濾膜過濾除菌。我把我用的protocol給您貼出來吧！ 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樓2010-05-16 20:21:30
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
|---|---|---|---|---|
|
[考研] 340求調劑 +6 | Amber00 2026-03-26 | 6/300 |
|
|---|---|---|---|---|
|
|
[考研] 085601材料工程找調劑 +9 | oatmealR 2026-03-29 | 9/450 |
|
|
|
[考研] 343求調劑 +6 | 愛羈絆 2026-03-29 | 6/300 |
|
|
[考研] 317分 一志愿南理工材料工程 本科湖工大 求調劑 +9 | 芋泥小鈴鐺 2026-03-28 | 9/450 |
|
|
|
[考研] 2026年華南師范大學歡迎化學，化工，生物，生醫(yī)工等專業(yè)優(yōu)秀學子加入！ +3 | llss0711 2026-03-28 | 6/300 |
|
|
|
[考研] 本科雙非材料，跨考一志愿華電085801電氣，283求調劑，任何專業(yè)都可以 +6 | 芝士雪baoo 2026-03-28 | 8/400 |
|
|
|
[考研] 0856求調劑 +13 | zhn03 2026-03-25 | 14/700 |
|
|
|
[考研] 289求調劑 +13 | 新時代材料 2026-03-27 | 13/650 |
|
|
|
[考研] 085600 286分 材料求調劑 +7 | 麻辣魷魚 2026-03-27 | 8/400 |
|
|
|
[考研] 286求調劑 +12 | PolarBear11 2026-03-26 | 12/600 |
|
|
|
[考研] 295求調劑 +5 | 1428151015 2026-03-27 | 6/300 |
|
|
|
[考研] 085701環(huán)境工程求調劑 +9 | 多久上課 2026-03-27 | 9/450 |
|
|
|
[考研] 265求調劑11408 +3 | 劉小鹿lu 2026-03-27 | 3/150 |
|
|
|
[考研] 292求調劑 +4 | 求求了收下我吧?/a> 2026-03-26 | 4/200 |
|
|
|
[考研] 【雙一流院校新能源、環(huán)境材料，材料加工與模擬招收大量調劑】 +4 | Higraduate 2026-03-22 | 8/400 |
|
|
|
[考研]
 085602 289分求調劑
+8
|
WWW西西弗斯 2026-03-24 | 8/400 |
|
|
|
[考研] 打過很多競賽，085406控制工程300分，求調劑 +3 | askeladz 2026-03-26 | 3/150 |
|
|
|
[考研] 07化學303求調劑 +5 | 睿08 2026-03-25 | 5/250 |
|
|
|
[考研] 材料專碩 335 分求調劑 +4 | 拒絕冷暴力 2026-03-25 | 4/200 |
|
|
|
[考研] 【2026考研調劑】制藥工程 284分 求相關專業(yè)調劑名額 +4 | 袁奐奐 2026-03-25 | 8/400 |
|