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kartiga natarajan

Member Since 19 Feb 2013
Offline Last Active May 13 2013 09:52 PM

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RNA biology

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2xzwei
12 Mar 2013 - 23:38
GNANA
20 Feb 2013 - 06:29

Posts I've Made

In Topic: quick change mutagenesis......

29 April 2013 - 02:59 AM

No the positive control is the maxi prep plasmid isolated from kit...But i screened the plasmids manually...Also i used Pfu Turbo DNA polymerase only...While isolating plasmids manually i did not give 70 percent ethanol wash...Will that be the reason?Im sure my competent cells are sterile...While running plasmids after isolation all plasmids showed same retardion compared to my empty vector. Will take your suggestions Gnana....Thanks

In Topic: quick change mutagenesis......

29 April 2013 - 01:30 AM

Gnana my gene was cloned in Xho1 & EcoR1 sites....So i took some plasmids from XL1 & DH5alpha transformed after following mutation protocol....But i could see out of them only one plasmid has released the insert...My positive control also have worked....What would be the reason? Shall i give the ones which have released the inserts after digestion 4 sequencing? I am also doing PCR with those plasmids...Kindly give me your valuable suggestions....

In Topic: quick change mutagenesis......

28 April 2013 - 10:01 PM

Hi Gnana...As per ur suggestions i prepared Top10 competent cells and i got colonies in both DH5alpha and XL1 blue competent cells....I also screened for the plasmids and the plasmids are also fine....further im also trying to verify using Restriction digestion and PCR...So give me an idea how many plasmids shall i send for sequencing? both from XL1 and DH5alpha? Thanks in advance

In Topic: quick change mutagenesis......

24 April 2013 - 07:21 AM

Is there any nice protocol to prepare this efficient competent cells other than Rbcl...Since in our lab it is not available....Can u give me a nice protocol?

In Topic: quick change mutagenesis......

24 April 2013 - 03:55 AM

Also after Dpn1 digestion i did not get any smear around the product as you said...I really dont know where will be the problem? plz do help me in sorting this Gnana...

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