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| 8 AUG 2003 at 2:49am |
AllanonJourneyman
Posts : 1322
Joined: 19 JUN 2003
Status : Offline | I'm free - kinda sucks sometime though !!
You only live once , but , if you live right , once is enough . Being happy takes work ...
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| 8 AUG 2003 at 2:51am |
| Deleted User | Hey come one Godfather... COme out of hiding and tell us now!
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| 8 AUG 2003 at 2:51am |
AllanonJourneyman
Posts : 1322
Joined: 19 JUN 2003
Status : Offline | Originally Posted By Jessica (7 AUG 2003 2:19pm)
Single in both worlds.
How could someone so beautiful be free ??
You only live once , but , if you live right , once is enough . Being happy takes work ...
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| 8 AUG 2003 at 2:53am |
AllanonJourneyman
Posts : 1322
Joined: 19 JUN 2003
Status : Offline | Originally Posted By Sarah (7 AUG 2003 5:11pm)
How about you Godfather? "What" are you?
And you , Sarah , what is your relationship status ??!! Cheers !
You only live once , but , if you live right , once is enough . Being happy takes work ...
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| 8 AUG 2003 at 4:25am |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Allanon (8 AUG 2003 2:51am)
How could someone so beautiful be free ?? I am asking myself the exact same thing :-/
I don't see why I should post such information before Godfather does Because I am asking... :
I have sometimes kids, some other times I am not married and I could also be a kid....or a baby with a really big head to store my brain... :
I FORGOT TO MENTION THAT IF I AM VERY YOUNG THEN MY PARENTS ALLOW ME TO STAY ON LINE FOR ABOUT 7 HOURS A DAY... :
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 4:27am |
| Deleted User | You're filled with mystery Godfather! :
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| 8 AUG 2003 at 4:40am |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By sennebec (7 AUG 2003 9:45pm)
GF... shame on you...
using this forum for a dating service : If you really don't like it -I do- we may start deleting our posts and make the thread dissapear....
I know you want some MORE action around here, right?
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 7:42am |
| Deleted User | I know you won't delete your posts. You're counting on them.
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| 8 AUG 2003 at 7:53am |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Sarah (8 AUG 2003 7:42am)
I know you won't delete your posts. You're counting on them. You are probably talking about my previous version...
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 8:48am |
| Deleted User | So, you've turned into a new leaf, eh?
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| 8 AUG 2003 at 9:22am |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Sarah (8 AUG 2003 8:48am)
So, you've turned into a new leaf, eh? Every night I die and every morning I am reborn. I am a new man everyday...
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 11:43am |
Agustín CordesGuild Master
Posts : 5696
Joined: 23 OCT 2002
Location: AR, Buenos Aires
Status : Offline | Originally Posted By Godfather (8 AUG 2003 4:25am)
I FORGOT TO MENTION THAT IF I AM VERY YOUNG THEN MY PARENTS ALLOW ME TO STAY ON LINE FOR ABOUT 7 HOURS A DAY... :
When I was a kid I stayed all day playing games (does it show?) and fooling around with the internet. Far more than 7 hours So that argument is basically nonsense
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| 8 AUG 2003 at 11:55am |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | No, I said that I am online 7 hours. My computer never shuts down! Movies, games, internet....my whole life....
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 12:03pm |
Agustín CordesGuild Master
Posts : 5696
Joined: 23 OCT 2002
Location: AR, Buenos Aires
Status : Offline | Originally Posted By Godfather (8 AUG 2003 11:55am)
No, I said that I am online 7 hours. My computer never shuts down! Movies, games, internet....my whole life....
Oh sure. I used to spend about 4 hours for chating, 3 hours for surfing and 6 hours for gaming.
So I still believe you're young
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| 8 AUG 2003 at 12:07pm |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Agustin (8 AUG 2003 12:03pm)
Oh sure. I used to spend about 4 hours for chating, 3 hours for surfing and 6 hours for gaming.
So I still believe you're young Your parents must have been away all day, right?
4+3+6=10
5 (sleeping)
6 (school)
3 (anything else)
You must have been a really busy boy.... :
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 12:11pm |
Agustín CordesGuild Master
Posts : 5696
Joined: 23 OCT 2002
Location: AR, Buenos Aires
Status : Offline | Yep, more or less. School was 5 hours though. And how about you? You say you spend 7 hours online plus movies plus games. Are you busy a boy?
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| 8 AUG 2003 at 12:21pm |
sennebecGuild Master
Posts : 3334
Joined: 15 NOV 2004
Location: US, maine
Status : Offline | Originally Posted By Godfather (8 AUG 2003 4:40am)
I know you want some MORE action around here, right?
???
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| 8 AUG 2003 at 12:22pm |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Agustin (8 AUG 2003 12:11pm)
Yep, more or less. School was 5 hours though. And how about you? You say you spend 7 hours online plus movies plus games. Are you busy a boy? Yeah, I guess that I am a very busy MAN...
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 12:25pm |
Agustín CordesGuild Master
Posts : 5696
Joined: 23 OCT 2002
Location: AR, Buenos Aires
Status : Offline | Originally Posted By Godfather (8 AUG 2003 12:22pm)
Yeah, I guess that I am a very busy MAN...
OK MAN, what do you do for a living then?
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| 8 AUG 2003 at 12:26pm |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By sennebec (8 AUG 2003 12:21pm)
???
You don't?!
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 12:29pm |
KsandraSchattenjger
Posts : 2459
Joined: 2 APR 2003
Status : Online | I know how old Godfather is... *hums annoyingly*
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| 8 AUG 2003 at 12:34pm |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | OK MAN, what do you do for a living then?
Reverse transcriptase-PCR (RT-PCR)--a method for assessing gene expression, detecting low copy number mRNA transcripts, and generating complementary DNAs (cDNAs) for cloning--converts an RNA template to its DNA counterpart with a reverse transcriptase enzyme. The cDNA is then amplified by PCR. Although the technique appears relatively painless, researchers using RT-PCR may encounter several pitfalls. For instance, reverse transcribing long mRNA templates or those with a high degree of secondary structure may be difficult with standard reverse transcriptases. The technique itself can be time consuming, because RNA extraction and purification is typically required before the reactions can take place. In addition, researchers who wish to use RT-PCR to quantitate steady-state levels of mRNA face challenges.
Fortunately, a number of companies have introduced products that overcome or mitigate many of the problems associated with RT-PCR. This article will review new developments in reverse transcriptases, kits for direct cDNA synthesis from cell lysates, and quantitative RT-PCR technologies.
The most commonly used reverse transcriptases are derived from avian myeloblastosis virus (AMV) or Moloney murine leukemia virus (MMLV). The accompanying table lists suppliers of MMLV-RT and AMV-RT as well as variants of these enzymes. Tth, a DNA polymerase that exhibits reverse transcriptase or DNA polymerase activity under different conditions and is also used for first-strand cDNA synthesis, has the advantage of higher thermal stability than AMV-RT or MMLV-RT. This increases the efficiency of reverse transcription by melting RNA secondary structure and decreases nonspecific primer binding. However, Tth is error prone, limiting its use.1,2 To solve this problem, Roche Molecular Biochemicals of Indianapolis offers C.therm. Polymerase, the Klenow fragment of the DNA polymerase from the thermophilic eubacterium Carboxydothermus hydrogenoformus. C.therm. Polymerase exhibits high thermal stability and offers high-fidelity first-strand cDNA synthesis, with a transcriptional error rate half that of Tth.
Recently several other companies have introduced new reverse transcriptase enzymes that offer high efficiency and/or improved synthesis of long cDNAs. Most of these products are standard reverse transcriptase enzymes with point mutations that enhance performance. To minimize degradation of RNA molecules during first-strand cDNA synthesis of long transcripts, companies have engineered reverse transcriptase variants with decreased RNase activity. CLONTECH of Palo Alto, Calif., offers the PowerScript™ Reverse Transcriptase, a point mutant of MMLV-RT that lacks RNase H activity but retains the polymerase activity of wild-type MMLV-RT to synthesize cDNAs of up to 3 kb. The PowerScript Reverse Transcriptase is guaranteed to be RNase- and DNase-free. Life Technologies Inc. of Rockville, Md., offers the SuperScript™ II RNase H- Reverse Transcriptase, an improved version of its SuperScript Reverse Transcriptase. The SuperScript II Reverse Transcriptase is also a point mutant of MMLV-RT that retains full polymerase activity. Promega Corp. of Madison, Wis., New England Biolabs of Beverly, Mass., Stratagene of La Jolla, Calif., and Epicentre of Madison, Wis., also offer RNase H-Minus variants of MMLV-RT.
-The Scientist
old trick...
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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| 8 AUG 2003 at 12:37pm |
Agustín CordesGuild Master
Posts : 5696
Joined: 23 OCT 2002
Location: AR, Buenos Aires
Status : Offline | And you expect me to believe that? How naive
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| 8 AUG 2003 at 12:41pm |
StammerGuild Master
Posts : 3894
Joined: 5 JAN 2003
Status : Online | Originally Posted By Agustin (8 AUG 2003 12:37pm)
And you expect me to believe that? How naive I have made you read it, didn't I
D@mn I am getting better at this day by day
Resistance is not futile, we're gonna win this thing, humankind is too good, we're not a bunch of under-achievers! We're gonna stand up, and we're gonna be human beings. We're going to get fired up about the real things, the things that matter! Creativity, and the dynamic human spirit that refuses to submit.
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