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實驗室手扎

Tuesday, May 22, 2007. Primary Culture of Mouse Dorsal root ganglian. Isolate mouse DRG and place into 15 ml DMEM (serum-free) solution. Centrifuge using 800rpm, at room temperature, for 2 minutes and then remove supernatant. Add 1 ml DMEM (serum-free) containing 0.125% Type Ia Collagenase (Stock=2.5%, take 50 ul into 950 ul DMEM), and incubate in 37 0C for 1.5 hours. Add 5 ml DMEM, and then gentled mix for a while. Centrifuge at 800rpm, RT for 2 minutes and then remove supernatant. Sunday, April 29, 2007.

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實驗室手扎 | shlin-labman.blogspot.com Reviews

<META> DESCRIPTION

Tuesday, May 22, 2007. Primary Culture of Mouse Dorsal root ganglian. Isolate mouse DRG and place into 15 ml DMEM (serum-free) solution. Centrifuge using 800rpm, at room temperature, for 2 minutes and then remove supernatant. Add 1 ml DMEM (serum-free) containing 0.125% Type Ia Collagenase (Stock=2.5%, take 50 ul into 950 ul DMEM), and incubate in 37 0C for 1.5 hours. Add 5 ml DMEM, and then gentled mix for a while. Centrifuge at 800rpm, RT for 2 minutes and then remove supernatant. Sunday, April 29, 2007.

<META> KEYWORDS

1 實驗室手扎

2 step 1

3 step 2

4 step 3

5 step 4

6 step 5

7 step 6

8 step 7

9 molecular cloning banding大量高純度的質體dna

10 滅好菌的lb培養液 500 ml/construct

CONTENT

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KEYWORDS ON PAGE

實驗室手扎,step 1,step 2,step 3,step 4,step 5,step 6,step 7,molecular cloning banding大量高純度的質體dna,滅好菌的lb培養液 500 ml/construct,測od值所需比色管,乾淨的紗布數塊,手提式紫外光燈,透析用dna袋子、夾子和裝4公升te溶液的容器，stir bar等,封口後invert數下，使etbr混合均勻，包覆錫泊紙以阻絕光線,上機前在確定兩試管重量相等，方可進行超高速離心,使用步驟如下,配方見下附註,製作方法

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實驗室手扎 | shlin-labman.blogspot.com Reviews

https://shlin-labman.blogspot.com

Tuesday, May 22, 2007. Primary Culture of Mouse Dorsal root ganglian. Isolate mouse DRG and place into 15 ml DMEM (serum-free) solution. Centrifuge using 800rpm, at room temperature, for 2 minutes and then remove supernatant. Add 1 ml DMEM (serum-free) containing 0.125% Type Ia Collagenase (Stock=2.5%, take 50 ul into 950 ul DMEM), and incubate in 37 0C for 1.5 hours. Add 5 ml DMEM, and then gentled mix for a while. Centrifuge at 800rpm, RT for 2 minutes and then remove supernatant. Sunday, April 29, 2007.

INTERNAL PAGES

shlin-labman.blogspot.com

1

實驗室手扎: October 2006

http://shlin-labman.blogspot.com/2006_10_01_archive.html

Tuesday, October 31, 2006. 在我們和生技公司購買大腸桿菌勝任細胞時，它通常是以零下-80度C的包裝運送為了保持這勝任細胞的活性，我們也應將原廠的原始菌種 stock 固定存放在-80度C的冷凍庫中。一首先，我們準備培養大腸桿菌的培養液LB broth 配方見下表一、保存勝任細胞的TSB緩衝液配方見下表二、預約好光譜儀 spectro-photometer 和低溫離心機 centrifugator。無菌取放3 ml的培養液在養菌專用的試管 Falcon 15 ml ，用乾淨的牙籤沾一點原始菌種加入試管，將試管放進37度C、240 rpm震盪的細菌培養箱培養16小時。二再準備50 ml LB培養液，倒入滅好菌的錐形瓶。將培養16小時的菌液取100 ul的量比例約1 500 加入，再放進細菌培養箱培養約3小時。三將菌液進行離心 Beckman Rotor ，離心的條件為4度C、3000 rpm歷時10分鐘，倒掉上方澄清的培養廢液，再用力敲打離心管使下方沈積的細菌塊鬆散，加入5 ml 置備50支勝任細胞的量 TSB緩衝液含5% DMSO。附帶一提，影響轉型...

2

實驗室手扎: molecular cloning 如何製作勝任細胞?

http://shlin-labman.blogspot.com/2006/10/molecular-cloning.html

Tuesday, October 31, 2006. 在我們和生技公司購買大腸桿菌勝任細胞時，它通常是以零下-80度C的包裝運送為了保持這勝任細胞的活性，我們也應將原廠的原始菌種 stock 固定存放在-80度C的冷凍庫中。一首先，我們準備培養大腸桿菌的培養液LB broth 配方見下表一、保存勝任細胞的TSB緩衝液配方見下表二、預約好光譜儀 spectro-photometer 和低溫離心機 centrifugator。無菌取放3 ml的培養液在養菌專用的試管 Falcon 15 ml ，用乾淨的牙籤沾一點原始菌種加入試管，將試管放進37度C、240 rpm震盪的細菌培養箱培養16小時。二再準備50 ml LB培養液，倒入滅好菌的錐形瓶。將培養16小時的菌液取100 ul的量比例約1 500 加入，再放進細菌培養箱培養約3小時。三將菌液進行離心 Beckman Rotor ，離心的條件為4度C、3000 rpm歷時10分鐘，倒掉上方澄清的培養廢液，再用力敲打離心管使下方沈積的細菌塊鬆散，加入5 ml 置備50支勝任細胞的量 TSB緩衝液含5% DMSO。附帶一提，影響轉型...

3

實驗室手扎: November 2006

http://shlin-labman.blogspot.com/2006_11_01_archive.html

Sunday, November 05, 2006. 所謂的banding是將質體DNA透過高密度介質氯化銫 CsCl 在超高速離心 ultracentrifuge 下形成分離，質體DNA在超高速離心的氯化銫中形成單一個帶狀故名。一第一天下午五點將單珠菌種移植到3 ml的LB培養液 A T 使用前加入tetracycline 12.5 ug/ml與ampicilin 50 ug/ml ，準備或預借之後所需的實驗材料如下. Beckman GSA 10 rotor離心用的250 ml離心管兩支 /construct 檢查每支管子的O-ring是否完好. Chloramphenicol 34 mg/ml in pure ethanol 2.5 ml/construct. Solution 1 30 ml /construct. Solution 2 60 ml /construct. Solution 3 45 ml /construct. Isopropanolol 0.6倍體積約81 ml /construct. 1xTE 溶解DNA用，pH=8.0、20 ml /construct. 五加入4...

4

實驗室手扎: April 2007

http://shlin-labman.blogspot.com/2007_04_01_archive.html

Sunday, April 29, 2007. Culture of mouse embryonic stem cell. 1) Mouse embryonic stem cell.medium preparation. 2) Pick-up stable targeting ES cell clone. After 5 7 days of culture with medium containing G418 and ganciclovir , ES cell showed surviving clones. The following protocol describes critical steps in picking up targeting ES clones and preparing genomic DNA from each clone. Drawing line with marker pen in the bottom of each 10-cm culture dish. Change Fresh ES medium 1 hour before picking up clones.

5

實驗室手扎: May 2007

http://shlin-labman.blogspot.com/2007_05_01_archive.html

Tuesday, May 22, 2007. Primary Culture of Mouse Dorsal root ganglian. Isolate mouse DRG and place into 15 ml DMEM (serum-free) solution. Centrifuge using 800rpm, at room temperature, for 2 minutes and then remove supernatant. Add 1 ml DMEM (serum-free) containing 0.125% Type Ia Collagenase (Stock=2.5%, take 50 ul into 950 ul DMEM), and incubate in 37 0C for 1.5 hours. Add 5 ml DMEM, and then gentled mix for a while. Centrifuge at 800rpm, RT for 2 minutes and then remove supernatant. 26519;星宏.

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shlin-labman.blogspot.com

實驗室手扎

Tuesday, May 22, 2007. Primary Culture of Mouse Dorsal root ganglian. Isolate mouse DRG and place into 15 ml DMEM (serum-free) solution. Centrifuge using 800rpm, at room temperature, for 2 minutes and then remove supernatant. Add 1 ml DMEM (serum-free) containing 0.125% Type Ia Collagenase (Stock=2.5%, take 50 ul into 950 ul DMEM), and incubate in 37 0C for 1.5 hours. Add 5 ml DMEM, and then gentled mix for a while. Centrifuge at 800rpm, RT for 2 minutes and then remove supernatant. Sunday, April 29, 2007.

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