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TheFFM
Member since: Dec 06, 2008
From: Ohio, United States
Status: Administrator & Neuroscience, Biophysics, and Microscopy Moderator
My points: 1353    what's this
Name: Fraser Moss
 


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Mar 19, 2010    10:55 AM

Replied to topic in Cell Culture and Tissue Culture forum   Mycoplasma in cell culture

There are many commercial mycoplasma removal agents available on the market. Here is one such product from AbD Serotec : http://www.abdserotec.com/antibodies/_-585.html The page includes a full protocol. Here are some alternatives http://www.invivogen.com/family.php?ID=3 http://www. ...

Mar 17, 2010    03:56 PM

Replied to topic in Neuroscience forum   Patch Clamp: Noise after making giga seal

Thanks for the update!

Mar 17, 2010    11:16 AM

Replied to topic in Cell Culture and Tissue Culture forum   Hep G2 cell culture

click on the linked title for the paper above for all their conditions...it's open access

Mar 17, 2010    10:43 AM

Replied to topic in Cell Culture and Tissue Culture forum   Hep G2 cell culture

This paper may be helpful BMC Syst Biol. 2007 May 11;1:21. A hierarchical approach employing metabolic and gene expression profiles to identify the pathways that confer cytotoxicity in HepG2 cells. Li Z, Srivastava S, Yang X, Mittal S, Norton P, Resau J, Haab B, Chan C. Department of Chemic ...

Mar 17, 2010    10:22 AM

Replied to topic in Cell Culture and Tissue Culture forum   Hep G2 cell culture

A culture media for those is RPMI-1640 containing: L-glutamine 2 mM fetal calf serum (10%) penicillin (100 U/ml) streptomycin (100 pg/ml) I have seen DMEM or MEM being used instead of RPMI too. The ATCC page says use MEM with 10% FBS http://www.atcc.org/ATCCAdvancedCatalogSearch/Produc ...

Mar 16, 2010    11:51 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   Sequencing Problems

Are you sequencing with primers that start in the vector and sequence through the cloned fragment? have you tried using primers within your predicted cloned fragment and that will sequence out into the vector at either end to see in which orientation the fragment has cloned? However before y ...

Mar 16, 2010    11:15 AM

Replied to topic in Anatomy and Physiology forum   problem with pore formation

With gramicidin concs of 0.1mg/ml I would expect that you will be seeing spontaneous rupturing of the membrane patch for most of your recordings. Try reducing your gramicidin to 20-25 µg/mL (1/4 of what you are presently using). And even then you must prefill the very tip of the electrod ...

Mar 16, 2010    09:38 AM

Replied to topic in Anatomy and Physiology forum   problem with pore formation

It sounds like your patch seal gets blown and/or the cell gets super leaky. What is the seal resistance after the holding current jumps to -3 nA? What concentration of gramicidin are you using? do you sonicate your pipette solution immediately before you backfill the pipette EVERY time ...

Mar 12, 2010    10:04 AM

Replied to topic in Anatomy and Physiology forum   Troubleshooting ACSF

Have you tried waiting until after you have bubbled the solution with 95% oxygen, 5% CO2 to saturation before you add the CaCl2?

Mar 11, 2010    03:26 PM

Variations include using these substitutes 1.8 mM CaCl2 (anhydrous) 199.8 mg/L OR 1.8ml/L 1M CaCl2 Stock solution 2ml/L 1M MgCl2 stock solution 667 ul/L of 3M KCL stock solution

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