Navigation Links
Mycobacterium intracellulare

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.521 02/2002 Microorganism Mycobacterium intracellulare 1403 Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium Middlebrook 7H9 liquid medium with oleic acid dextrose complex (OADC) Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Middlebrook 7H9 liquid medium with oleic acid dextrose complex Cuvette 2 mm gap width Reference Marklund, B.-I. et al 1995 Journal of Bacteriology 177, No. 21 6100-6105 Making electrocompetent cells:

1. Grow a 50-100 ml cell culture at 37 C statically to an O.D.600 of 0.1 to 0.3. 2. Harvest by centrifugation. 3. Wash twice in cold 10% glycerol. The first time in 30 ml the second time in 1 ml. 4. Resuspend 0.2-0.5 ml of 10% glycerol.

Electroporation of cells:< /b>

  1. Add 1 g plasmid DNA to 100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into a prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,500 V Time constant (T) 5 ms
  4. Transfer cells to 1 ml of 7H9 medium and grow statically for 16-20 h.
  5. To disperse cells, bath sonicate cultures twice for 30 s each time. Dilute suspension in 0.9% NaCl supplemented with 0.1% Tween 80.
  6. Plate on selective agar.
Expected Results: Transformation efficiency up to 106 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Mycobacterium avium
2. Mycobacterium smegmatis
3. Mycobacterium tuberculosis
4. Mycobacterium avium
5. Mycobacterium intracellulare
6. Mycobacterium smegmatis
7. Mycobacterium tuberculosis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/3/2016)... ... February 03, 2016 , ... ... annual report which summarizes and analyzes nearly 750 unique supply chain notifications ... and analysis service. , Supply chain risk management practitioners subscribe to the ...
(Date:2/3/2016)... , Feb. 3, 2016  Today, Symphony Technology ... AlphaImpactRx , a leading provider of primary research ... to IMS Health , a global information and ... offerings, capabilities and technologies will be integrated into IMS ... global primary market research capabilities. ...
(Date:2/3/2016)... MN (PRWEB) , ... February 03, 2016 , ... ... physiologic monitoring, has released two new solutions focused on social housing of small ... and reduces stress artifacts in data.¹, ² , DSI’s HD-S11 implant has evolved ...
(Date:2/3/2016)... , ... February 03, 2016 , ... Big games come ... have sacrificed so much for our country. CereScan, a nationally recognized brain diagnostics and ... Wounded Warrior Amputee Football Team (WWAFT) vs. NFL Stars Flag Football Game on February ...
Breaking Biology Technology:
(Date:2/2/2016)... MOUNTAIN VIEW, Calif. , Feb. 2, 2016 ... diabetic retinopathy market, Frost & Sullivan recognizes US-based ... North America Frost & Sullivan Award for New ... technology provider in North America ... standard in the rapidly growing diabetic retinopathy market. ...
(Date:2/1/2016)... 1, 2016  Wocket® smart wallet ( www.wocketwallet.com ) announces the launch ... Joey Fatone . Las Vegas , where Joey ... --> Las Vegas , where Joey appeared at the ... new video ad was filmed at the Consumer Electronics Show (CES2016) in ... booth to meet and greet fans. --> ...
(Date:1/25/2016)... BLUE BELL, Pa. , Jan. 25, 2016   Unisys ... facial recognition system at John F. Kennedy (JFK) International Airport, ... and Border Protection (CBP) identify imposters attempting to enter ... or do not belong to them. pilot testing ... rolled out initially at three terminals at JFK during January 2016. ...
Breaking Biology News(10 mins):