 |
 |
 |
 |
 |
 |
||
 |
 |
 |
 |
 |
| 5 | 1/1 | 返回列表 |
| 查看: 1409 | 回復(fù): 5 | |||
| 當(dāng)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 | |||
粉色水晶新蟲 (初入文壇)
|
[求助]
乳酸菌電轉(zhuǎn) 已有1人參與
|
||
|
求助各位大神有沒有知道乳酸乳球菌MG1363感受態(tài)的具體做法及原理，和電轉(zhuǎn)方法及原理！謝謝！ @youlinglyw @wizardfan @biostar2009 發(fā)自小木蟲Android客戶端 |
回復(fù)此樓» 猜你喜歡
不合理蛙科研實驗中的趣事：實驗器材的 “烏龍”
已經(jīng)有0人回復(fù)
-
不合理蛙科研實驗中的趣事：和實驗材料的 “斗智斗勇”
已經(jīng)有0人回復(fù)
-
化學(xué)工程及工業(yè)化學(xué)論文潤色/翻譯怎么收費?
已經(jīng)有219人回復(fù)
-
蛋白質(zhì)檢測：精準(zhǔn)分析，解鎖生物分子的密碼
已經(jīng)有0人回復(fù)
-
不合理蛙科研實驗之小鼠實驗：嚴(yán)謹(jǐn)設(shè)計，解析生命機制的重要載體
已經(jīng)有0人回復(fù)
-
不合理蛙科研實驗之重金屬檢測：精準(zhǔn)篩查，守護健康與環(huán)境的防線
已經(jīng)有0人回復(fù)
-
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(fù)
-
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(fù)
-
納米粒度分析
已經(jīng)有3人回復(fù)
-
林科院林化所招收材料與化工專業(yè)碩士，坐標(biāo)南京
已經(jīng)有0人回復(fù)
WilliamHJ
金蟲 (小有名氣)
- 應(yīng)助: 8 (幼兒園)
- 金幣: 2186.9
- 紅花: 5
- 帖子: 170
- 在線: 58.5小時
- 蟲號: 1858225
- 注冊: 2012-06-13
- 性別: GG
- 專業(yè): 生物物理、生物化學(xué)與分子
【答案】應(yīng)助回帖
★ ★ ★
西門吹雪170: 金幣+3, 鼓勵回帖交流 2020-03-11 08:06:52
西門吹雪170: 金幣+3, 鼓勵回帖交流 2020-03-11 08:06:52
|
Preparation of the cells: Day 1: Inoculate 5 ml of G/L-SGM17B medium with L. lactis glycerol stock from -80 C and grow at 30 C, without aeration, overnight Day 2: Inoculate 50 ml of G/L-SGM17B with pre-culture in a dilution of 1:100 and grow at 30 C, without aeration, overnight Day 3: - Add 50 ml full-grown culture to 400 ml of G/L-SGM17B medium - Grow the culture until OD600 is 0.2-0.3 (ca. 3 h) - Spin down cells for 20 min at 6000 x g, 4 C - Wash cells with 400 ml of 0.5 M sucrose, 10% glycerol (4 C) and spin down at 6000 x g (centrifugation speed may need to be increased during successive washing steps) - Resuspend the cells in 200 ml of 0.5 M sucrose, 10% glycerol, 50 mM EDTA (4 C), keep the suspension on ice for 15 min and spin down - Wash cells with 100 ml of 0.5 M sucrose, 10% glycerol (4 C) and spin down (6000 x g) - Resuspend the cells in 4 ml of 0.5 M sucrose, 10% glycerol (4 C): Use 40 μl per electroporation (keep on ice) Or store the cells in small portions at -80 C, let them defreeze on ice before use Electroporation: - Place 40 μl cells in a pre-chilled electroporation cuvette with 1 μl DNA (100-500 ng vector DNA reconstituted in TE-, Tris-buffer, or distilled water; for transforming cells with ligation product use 500-1000 ng DNA) and keep the cuvette on ice - Use Biorad Genepulser with following adjustments: 2000 V 25 μF 200 Ω - Pulse (normal reading is 4.5-5 msec) - Add 1 ml of G/L-M17B + 20 mM MgCl2 + 2 mM CaCl2 - Keep the cuvette for 5 min on ice and incubate 1-1.5 h at 30 C - Plate 10 μl, 100 μl, 900 μl on M17agar with glucose or lactose and antibiotics (depends on plasmid) - Incubate 1-2 days at 30 °C Materials: - G/L-SGM17B: M17-Broth with: 0.5 M sucrose 2.5% glycine 0.5% glucose or 0.5% lactose (strain dependent) Add the sucrose and glycine to the M17-B and sterilize 20 min 121 °C. Add sterile glucose or lactose after cooling down. - 0.5 M sucrose/ 10% glycerol - 0.5 M sucrose/ 10% glycerol/ 0.05 M EDTA L. lactis grows very slowly on G/L-SGM17B. Leaving out the sucrose is possible (Wells et al., 1993) but can decrease the transformation efficiency. The medium for cell recovery must contain MgCl2 and CaCl2. 我用這個protocol做的，效果還不錯 |
5樓2020-03-10 11:17:16
2樓2019-06-16 11:15:32
rabbitnxm
新蟲 (初入文壇)
- 應(yīng)助: 0 (幼兒園)
- 金幣: 23.3
- 帖子: 20
- 在線: 1.7小時
- 蟲號: 14891109
- 注冊: 2019-04-16
- 專業(yè): 微生物生理與生物化學(xué)
3樓2019-08-09 22:34:26
rabbitnxm
新蟲 (初入文壇)
- 應(yīng)助: 0 (幼兒園)
- 金幣: 23.3
- 帖子: 20
- 在線: 1.7小時
- 蟲號: 14891109
- 注冊: 2019-04-16
- 專業(yè): 微生物生理與生物化學(xué)
4樓2019-08-09 22:36:05
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
|---|---|---|---|---|
|
[考研] 材料與化工（0856）304求B區(qū)調(diào)劑 +6 | 邱gl 2026-03-10 | 9/450 |
|
|---|---|---|---|---|
|
|
[考研] 274環(huán)境工程求調(diào)劑 +8 | 扶柳盈江 2026-03-05 | 9/450 |
|
|
|
[考研] 308求調(diào)劑 +3 | 是Lupa啊 2026-03-11 | 3/150 |
|
|
[考研] 材料工程085601調(diào)劑求老師收留 +8 | 強木木木 2026-03-07 | 10/500 |
|
|
|
[考研] 312求調(diào)劑 +5 | 陌宸希 2026-03-10 | 5/250 |
|
|
|
[考研] 420求調(diào)劑 +3 | 莫向外求11 2026-03-10 | 3/150 |
|
|
|
[考研] 調(diào)劑 +5 | 呵唔哦豁 2026-03-10 | 5/250 |
|
|
|
[考研] 308求調(diào)劑 +4 | 是Lupa啊 2026-03-08 | 7/350 |
|
|
|
[考研]
 調(diào)劑的同學(xué)，走過路過，不要錯過
+11
|
likeihood 2026-03-06 | 16/800 |
|
|
|
[考研] 313求調(diào)劑 +4 | Yyt楊1 2026-03-07 | 5/250 |
|
|
|
[考研] 材料調(diào)劑 +4 | xxxcm 2026-03-08 | 7/350 |
|
|
|
[考研] 0703化學(xué)求調(diào)劑，總分309分，一志愿華南師范 +3 | 花與葉@ 2026-03-08 | 3/150 |
|
|
|
[考研] 0856材料與化工290求調(diào)劑 +7 | Nebulala 2026-03-08 | 8/400 |
|
|
|
[考研] 083000環(huán)境科學(xué)與工程調(diào)劑 +5 | 加油呀fxy 2026-03-07 | 6/300 |
|
|
|
[考研]
 085600材料與化工 292分 求調(diào)劑
+6
|
程晴之 2026-03-06 | 6/300 |
|
|
|
[考研] 求調(diào)劑 +4 | 呼呼？~+123456 2026-03-05 | 5/250 |
|
|
|
[考研]
材料工程調(diào)劑
+6
|
zbcm_zbcm 2026-03-05 | 6/300 |
|
|
|
[考研] 化工282求調(diào)劑一志愿211 +5 | NA0912 2026-03-05 | 6/300 |
|
|
|
[考研] 304求調(diào)劑 +3 | 曼殊2266 2026-03-04 | 3/150 |
|
|
|
[考研] 293求調(diào)劑 +3 | 是樂渝哇 2026-03-04 | 3/150 |
|