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unguyan
Member since: Mar 18, 2007
From: Jiangsu, China
Status: Frog Egg
My points: 41    what's this
Name: [Privacy]
 


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May 11, 2011    01:27 AM

Posted comment on protocol   simple method for DNA extraction of geram + bacteri

heat upto 100? for 10min

May 11, 2011    01:27 AM

Posted comment on protocol   simple method for DNA extraction of geram + bacteri

heat upto 100? for 10min

Jun 17, 2009    04:55 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[b]The experiment won through finally! All the problems is because of the template problem. And my experiment has proved that having smear is normal, in contrast, no smear is incredible. Thanks all of you![/b]

Apr 30, 2009    08:34 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote]parvoman : 4/28/2009 10:30 AM[/quote] You may would like to read this paper "[url=http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WFH-4TW14W7-4&_user=1003024&_coverDate=02%2F28%2F2009&_alid=910681218&_rdoc=1&_fmt=high&_orig=search&_cdi=6795&_sort=d&_st=4&_docanchor=&_ct=1&_acct= ...

Apr 28, 2009    11:14 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote=parvoman]Your three results suggest that the smear is due to genomic DNA priming / extension. ie. bits of genomic DNA anneal to other bits and the polymerase extends them. (To get the smear you need the genomic DNA and the polymerase but not the primers)[/quote] so, what's your seggust to ...

Apr 24, 2009    10:53 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote=parvoman] Did you get a chance to try out your positive control template in increasing amounts of non-infected rabbit serum-derived template DNA? If you find that a small amount of rabbit genomic DNA masks your positive signal then it means that you have to change your detection system. ...

Apr 20, 2009    01:48 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote=MegCD] What do the abbreviations at the top of your lanes stand for (1w, 2w, 3w etc). It seems like the band tapered off, though what I think is your positive control looks great! [/quote] "w" is a abbreviation of week. It means the rabbit infection time. So, the band (1w to 7w ...

Apr 19, 2009    03:36 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote] Yep, and we hope that you'll solve it soon PS. MegCD's point about using a shorter extension time is important - it should make your PCR more specific for the short product you're looking for. [/quote] 95°5 min 95°30sec 62°30sec 72°30sec 35cycles 72°5 ...

Apr 16, 2009    08:45 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote]Yep, and we hope that you'll solve it soon PS. MegCD's point about using a shorter extension time is important - it should make your PCR more specific for the short product you're looking for. [/quote] My Taq may have some problems. I am waiting for the new Taq. It may take some days ...

Apr 14, 2009    09:21 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   smear problem(have a picture of the gel)

[quote=MegCD] I'm surprised about the length of your cycling times - do you normally use these conditions? I typically use: 1. 95C for 5 min 2. 95C for 30 sec XXC for 30 sec 72C for 30 sec (1 min per 1 kb of product, usually don't go below 30 sec even if my product is shorter than 50 ...

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