 |
 |
 |
 |
 |
 |
||
 |
 |
 |
 |
 |
| 6 | 1/1 | 返回列表 |
| 查看: 1385 | 回復(fù): 5 | ||
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157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(jī)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(hù)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)
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
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
王超福瑞達(dá)
新蟲 (初入文壇)
- 應(yīng)助: 0 (幼兒園)
- 金幣: 153.5
- 帖子: 6
- 在線: 10.2小時
- 蟲號: 20380045
- 注冊: 2020-01-03
- 專業(yè): 生物大分子結(jié)構(gòu)與功能
6樓2020-04-10 15:37:30
相關(guān)版塊跳轉(zhuǎn)
我要訂閱樓主 粉色水晶 的主題更新
| 6 | 1/1 | 返回列表 |
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
|---|---|---|---|---|
|
[考研] A區(qū)一本交叉課題組，低分調(diào)劑，招收機(jī)械電子信息通信等交叉方向 +11 | lisimayy 2026-03-04 | 19/950 |
|
|---|---|---|---|---|
|
[考研] 334求調(diào)劑 +3 | Trying] 2026-03-05 | 4/200 |
|
|
|
[考研] 313求調(diào)劑 +3 | 水流年lc 2026-02-28 | 4/200 |
|
|
|
[考研] 304求調(diào)劑 +3 | 曼殊2266 2026-03-04 | 3/150 |
|
|
|
[考研] 0703化學(xué) 學(xué)碩 理工科均可 不區(qū)分研究方向 總分279求調(diào)劑 +7 | 1一11 2026-03-03 | 7/350 |
|
|
|
[考研] 304求調(diào)劑 +7 | 曼殊2266 2026-02-28 | 8/400 |
|
|
|
[考研] 復(fù)試調(diào)劑 +3 | 呼呼？~+123456 2026-03-05 | 5/250 |
|
|
|
[論文投稿]
 主編終審卡者不動
 100+4
|
Stray2021 2026-03-03 | 4/200 |
|
|
|
[考研] 307求調(diào)劑 +4 | xuyuan111 2026-03-04 | 5/250 |
|
|
|
[考研] 武理材料工程302調(diào)劑 +3 | Doleres 2026-03-03 | 6/300 |
|
|
|
[考研] 312求調(diào)劑 +8 | 醋精華了一下發(fā)?/a> 2026-03-03 | 10/500 |
|
|
|
[考研] 331求調(diào)劑 +3 | zzZ&zZ 2026-03-03 | 3/150 |
|
|
|
[考研] 293求調(diào)劑 +4 | 是樂渝哇 2026-03-03 | 4/200 |
|
|
|
[考研] 291求調(diào)劑 +4 | Afy123456 2026-03-03 | 7/350 |
|
|
|
[考研] 268求調(diào)劑 +10 | 簡單點0 2026-03-02 | 14/700 |
|
|
|
[考研] 材料學(xué)碩318求調(diào)劑 +11 | February_Feb 2026-03-01 | 11/550 |
|
|
|
[考研] 295求調(diào)劑。一志愿報考鄭州大學(xué)化學(xué)工藝學(xué)碩，總分295分 +8 | yl1 2026-03-02 | 9/450 |
|
|
|
[考研] 一志愿中石油（華東）本科齊魯工業(yè)大學(xué) +3 | 石能偉 2026-03-02 | 3/150 |
|
|
|
[考研] 哈工大計算機(jī)劉劼團(tuán)隊招生 +4 | hit_aiot 2026-03-01 | 6/300 |
|
|
|
[考研] 0856材料求調(diào)劑 +4 | 麻辣魷魚 2026-02-28 | 4/200 |
|