2008-2009 CAMPAIGN

April 19th, 2008 by nwoclone-army

REALM DIVISION 2008-2009

KenneapolisMaskirovka Fawkes FarmRongelo Island

Gliese-581Delta City,KenBalabac IslandVice City-Ken

Brgy.GinebraMischief ReefWashington,Ken

D’ArthayneianOrang MedanAjima Island

SamontesonianBubble-IslandEdo City

Realm City,KenRealmcity-kenKennex-Ken

Lusong GuoMarbbadenKapitbahayan

DonnanNWO RealmLiwayway

ReginaldOtaka-MaruLaong-Laan

Luzon EmpireGunkanjimaDimasalang

CastlebergNWO ManilaKagitingan

Quiet LodgeCapitol HillsManila,Ken

Sabah-RPHoly City,CATimes Beach

Ken EmpireAlphine VillageMoonwalker,Ken

Fort FoureauIsland Estate Racccoon-City

StauffenbergHorobinaiFuga Island

Mactan IslandSeattle-KenSulu Empire

Corregidor IslandAmbrose-KenLos Colonias

The Tank ManTambobongAgravante City

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April 4th, 2008 by nwoclone-army

RheoSwitch® Mammalian Inducible Expression System
Catalog # Size Concentration Price Qty
E3000S     $650.00
Prices are in US dollars and valid only for US orders.
Download: Technical Bulletin|Manual|MSDS PDF


  • Precise regulation of expression levels
  • Negligible basal expression
  • Synthetic inducer and engineered receptor eliminate non-specific side effects
  • Greater than 1,000 fold induction levels routinely obtained
  • No special culture and media requirements

Description:
The RheoSwitch® Mammalian Inducible Expression System permits maximum control of gene expression in mammalian cells. Analogous to the operation of a rheostat, the RheoSwitch technology allows induction and adjustable control of gene expression. This precise regulation of gene expression is achieved through the highly specific interaction of a synthetic inducer, RheoSwitch Ligand RSL1, and a chimeric bipartite nuclear receptor. This receptor is activated in the presence of RSL1 ligand, and the level of gene expression can be regulated by adjusting the concentration of RSL1 ligand in the tissue culture media.

The RheoSwitch technology offers several advantages over other inducible systems. Its precise control is unrivaled among mammalian expression systems, giving negligible levels of basal expression in the absence of inducer and greater than 10,000 fold induction when RSL1 ligand is present. Unlike other systems, which rely on steroids or other drugs, the synthetic ligand RSL1 shows no pleiotropic effects in mammalian cells and exhibits no cross talk with endogenous transcription factors. In addition, the RheoSwitch System has no special culture media requirements.





Figure 1: Chemical structure of RheoSwitch Ligand RSL1 (MW=382.5 daltons)





Figure 2: Transcriptional control using the RheoSwitch System. "OFF" state: In the absence of RSL1 ligand, the RheoReceptor and RheoActivator proteins exist in an inactive conformation and transcription is kept off. "ON" State: In the presence of RSL1 ligand, the two proteins stably dimerize and bind to the response element of pNEBR-X1Hygro in an active conformation and transcription is turned on.





Figure 3: Luciferase expression in response to increasing RSL1 concentrations. NIH 3T3 cells co-transfected with pNEBR-R1 and pNEBR-X1 expressing firefly luciferase were induced with the indicated RSL1 concentrations. Expression was measured 48 hours post-induction.





Figure 4: NIH 3T3 (A) and RheoSwitch HEK293-A7 (B) (NEB #C2003) cells were transfected with pNEBR-X1Hygro encoding a HA-tagged protein using TransPass D2 (NEB #M2554). NIH 3T3 cells were also co-transfected with pNEBR-R1. RSL1 was added to transfected cells 16 hours later, and cells were harvested 24 hours after induction (Note: DMSO is added as a control for 0 nM RSL1). Harvested cells were lysed, and equivalent amounts of protein (as determined by Coomassie Stain) were electrophoresed and transferred to nitrocellulose. An immunoblot assay was performed on the filter using anti-HA as primary antibody and detected using anti-mouse secondary antibody and reagents. A longer exposure of the HEK293-A7 immunoblot is shown (C) to illustrate that no protein is detected in the absence of RSL1.





Figure 5: Induction time course. NIH 3T3 cells co-transfected with pNEBR-R1 and pNEBR-X1GLuc control plasmid were induced with RSL1. Gaussia luciferase activity was assayed between 1 and 24 hours post-induction.





Figure 6: Rapid ON:OFF switch response to two cycles of addition and removal of RSL1. NIH 3T3 cells were co-transfected with pNEBR-R1 and pNEBR-X1 expressing a destabilized firefly luciferase and induced with RSL1 followed by removal of RSL1 at 24 hours. A second cycle of addition/removal of RSL1 results in a new ON:OFF switch response. To remove RSL1 and turn off expression, cells were washed and RSL1-free media was added at the times indicated. The blue line represents induced expression. The red line represents uninduced expression.



Background:
The RheoSwitch System represents the next generation of inducible gene expression systems for mammalian cells. It is composed of an engineered nuclear receptor and a synthetic ligand inducer that is highly specific. The system is built on a two-hybrid switch format that activates transcription in the presence of inducer and represses gene expression in its absence (1,2,3 and unpublished observations). The synthetic receptor is composed of two proteins, RheoReceptor-1 and RheoActivator, that dimerize to make a holoreceptor. Both are expressed from strong constitutive promoters on plasmid pNEBR-R1. The RheoReceptor-1 protein is a highly engineered ligand-binding domain (LBD) of an insect EcR nuclear receptor fused to the yeast GAL4 DNA binding domain. The RheoActivator protein is an insect/mammalian RXR hybrid LBD fused to the viral activation domain VP16. The gene to be expressed is cloned into the pNEBR-X1 plasmid under control of five tandem repeats of the GAL4 response element (5XRE). In the absence of RSL1 ligand, the receptor represses transcription by binding to the GAL4 elements in a transcriptionally inactive conformation. Upon induction, the RSL1 ligand tightly binds and changes the conformation of the RheoReceptor-1 protein which stabilizes the holoreceptor heterodimer on the 5XRE. The activated holoreceptor and the VP16 activation domain bind and recruit to the promoter transcriptional coactivators along with basal machinery, resulting in a highly induced transcriptional state (Figure 2).

This high induction of transcription coupled with extremely low basal expression results in extremely high induction levels (greater than 10,000 fold induction), with a degree of control that is superior to other systems.

RSL1 ligand induces transcription over a broad range of concentrations from picomolar to micromolar. As illustrated in Figure 3, Gaussia Luciferase activity increases in an almost linear fashion with increasing amounts of RSL1. Likewise, increasing amounts of protein are observed, via immunoblot assay, with increasing amounts of RSL1 (Figure 4). Even with a longer exposure, no protein is observed in non-induced cells (Figure 4, Lane 5), further demonstrating the degree of control of over expression. Induced expression can be seen in as little as 1 hour, reaching a maximum level between 48 and 72 hours (Figure 5). The RheoSwitch System is very responsive to RSL1 and its removal from the medium will rapidly turn off gene expression (Figure 6).

RSL1 ligand is a synthetic diacylhydrazine, [N-(2-ethyl-3-methoxybenzoyl)-N´-(3,5-dimethylbenzoyl)-N´-tert-butylhydrazine] (Figure 1) (4). It is one of a family of compounds that have been found to act as non-steroidal ecdysone agonists and can function as gene inducers (4,5). RSL1 binds tightly and selectively to RheoReceptor-1, altering its conformation so that it releases bound negative regulatory cofactors and stabilizes the RheoReceptor/RheoActivator heterodimer. RSL1 is not a steroid,it is invisible to mammalian nuclear receptors and has been shown to be inert within all cell lines tested to date. The result is an inducible system that specifically controls the gene of interest without unwanted effects on the host cells.

Applications:

  • Inducible protein expression
  • Expression of toxic genes
  • Overexpression/mutant rescue studies

Kit Components:
Gaussia Luciferase Assay Kit
pNEBR-R1 Regulator Plasmid (500 μg/ml)
pNEBR-X1GLuc Control Plasmid (500 μg/ml)
pNEBR-X1Hygro Vector
RheoSwitch® Ligand RSL1
GLuc Assay Buffer
GLuc Substrate (100X)
RheoSwitch® R-X1 Sequencing Primer (17-mer) (200 pmol)


Storage Conditions


Storage Temperature:
-20°C


FAQs

  1. Where can I find more detailed FAQs for RheoSwitch® Mammalian Inducible Expression System?
  2. What cell lines and systems can be used with RheoSwitch?
  3. Can RheoSwitch be used in mice?
  4. Is RSL1 stable at room temperature?
  5. Can RSL1 be diluted in media?
  6. I have cloned my gene into pNEBR-X1. Can I make a stable cell line with this plasmid?
  7. How can I assay for expression of the RheoReceptor and RheoActivator genes?
  8. Can I make a stably transformed cell line using pNEBR-R1?
  9. What concentration of G418 should I use to construct a stable cell line with pNEBR-R1?



Figure 6A: pNEBR-X1Hygro plasmid map. Used to clone the gene of interest, it has five copies of the GAL4 response element (5XRE) upstream of a TATA box and a short leader sequence followed by multiple cloning sites and an SV40 polyA signal. Only unique restriction enzyme sites are shown.





Figure 6B: pNEBR-X1Hygro multiple cloning sites (MCS). To minimize uninduced expression, a short sequence containing the TATA box separates the end of the 5XRE from the predicted transcription start. Only unique restriction enzyme sites are shown.





Figure 7: pNEBR-R1 plasmid map. Expresses an engineered nuclear receptor heterodimer, consisting of two proteins, RheoReceptor-1, and RheoActivator. The two proteins constitute the holoreceptor and regulate transcription of genes cloned into the expression vector, pNEBR-X1. The two proteins are expressed from the constitutive UbC and UbB promoters, respectively. The plasmid also contains the neomycin resistance gene under control of the SV40 early promoter for the generation of stable cell lines.


References

  1. Palli, S.R., Kapitskaya, M.Z., Kumar, M.B. and Cress, D.E. (2003) Eur. J. Biochem., 270, 1308-1315.
  2. Dhadialla, T.S., Carlson, G.R. and Le, D.P. (1998) Annu. Rev. Entomol., 43, 545-569.
  3. Kumar, M.B., Potter, D.W., Hormann, R.E., Edwards, A., Tice, C.M., Smith, H.C., Dipietro, M.A., Polley, M., Lawless, M., Wolohan, P.R., Kethidi, D.R. and Palli, S.R. (2004) J. Biol. Chem., 279, 27211-27218.
  4. Verhaegent, M. and Christopoulos, T.K. (2002) Anal. Chem., 74, 4378-4385.

Reagents Sold Separately


Gaussia Luciferase Assay Kit
pNEBR-R1 Regulator Plasmid
pNEBR-X1GLuc Control Plasmid
pNEBR-X1Hygro Vector
RheoSwitch® Ligand RSL1
RheoSwitch® R-X1 Sequencing Primer (17-mer)


Companion Products


pNEBR-X1 Vector
RheoSwitch® Cell Line CHO-R5
RheoSwitch® Cell Line CHO-R5 and pNEBR-X1GLuc Control Plasmid
RheoSwitch® Cell Line HEK293-A7
RheoSwitch® Cell Line HEK293-A7 and pNEBR-X1GLuc Control Plasmid
RheoSwitch® Cell Line NIH3T3-47
RheoSwitch® Cell Line NIH3T3-47 and pNEBR-X1GLuc Control Plasmid
TransPass™ D1 Transfection Reagent
TransPass™ D2 Transfection Reagent

Legal


Licenses/Patents/Disclaimers:
RheoSwitch® Mammalian Inducible Expression System Notice to Purchaser: This product is covered by patents owned or licensed by RheoGene, Inc. and licensed to New England Biolabs, Inc. The purchase of this product conveys to the non-commercial purchaser the non-transferable right to use the purchased amount of the product and components of the product for research use only conducted by the purchaser. For commercial purchasers using this product for research, the purchase of this product converys to the purchaser the non-transferable right to use the purchased amount of the product and components of the product for a six (6) month evaluation period after the date of purchase for research use only. After the six (6) month evaluation period, the purchaser must obtain a commercial research license from RheoGene, Inc. in order to continue use of the RheoSwitch® System.

pNEBR-X1-GLuc Gaussia Luciferase Control Plasmid Non-Commercial entities: This product is covered by US Patents # 6,232,107 and other patents that are the legal property as assigned to Prolume Ltd./Nanolight Technologies. This product is licensed only to the purchasing laboratory-research group. Recipient agrees not to transfer this plasmid or derivatives of this vector to any other laboratory, person or research group, even if within the same institution. Recipient agrees not to alter or make any changes to the nucleotide coding sequence or secretory coding sequence of the luciferase(s) contained within without prior written permission from Prolume Ltd./Nanolight Technologies (www.nanolight.com). Recipient agrees not to file for any patent rights or to any inventions claiming any portion of the luciferase(s) within this material without prior written permission from Prolume Ltd./Nanolight Technologies.

pNEBR-X1GLuc Gaussia Luciferase Control Plasmid Commercial For-Profit Entities & Non-Profit Foundations (herein referred to as Commercial Recipients): Commercial Recipients wishing to derive products, engage in the sale or license of any products, discover drugs, or make inventions by use of the materials enclosed, fully agree to the terms mentioned above for Non-Commercial entties; AND ADDITIONALLY agree to an are hereby bound to use the materials FOR EVALUATION PURPOSES ONLY. Commercial Recipient hereby agrees to destroy and cease use of any materials or derivatives containing any portion of these materials within 120 days from receipt. Commercial Recipient agrees not to use the materials for any use, other than the 120 day suitability evaluation without prior written permission or obtaining a valid license from Prolume Ltd./Nanolight Technologies

Privacy, Limitations, Warranty, Disclaimer, Copyright & Trademark

Dating Tips 101

April 2nd, 2008 by nwoclone-army

Be yourself. Better yet, be the best version of yourself when meeting other people. That way you don’t disappoint others and you don’t frustrate yourself.
Preening and Pruning practices. Translated as good grooming for guys and primping for girls. In dates, personal hygiene is of utmost importance. You won’t get a second chance on a first impression. So make sure you leave a LONG LASTING fresh and positive impression.
Time is of the essence. So be on time whenever, wherever, however you are meeting up. For guys this means being with their dates on time, and, for girls – zero waiting time for guys.
Interestingly enough. Be interested and interesting. Spark some interest. Ask appropriate and relevant questions. The point is you’ll never get bored if you, yourself, are not boring.
Reality Check. Focus all energies on your date and never check other people out. Subtlety has no place in an environment where you’re on a microscope. Extend the courtesy of concentrating solely on your date when you are with them.
Talk Now. Text Later. When dating, do what daters do: Talk. Keep cell phones ex communicated until the end of the date. You want to get to know each other by actually talking to each other – not others.
Laugh a little. Pack that sense of humor. It’s a date not a summit convention! So open up and share short anecdotes on topics you both can easily relate to.
Sense and sensibilities. Mind your manners. You are opening yourself out to the world but be mindful of socially accepted behavior. Respect should be high up on both you and your date’s list.
Expiration date. If you notice you have some issues with drinking in moderation, it’s best to stay away from something that will jeopardize your date. You don’t want to make a fool of yourself while you’re sober so why hurt your chances by getting drunk?
Ever after. Make sure to thank your date for the experience; regardless of the possibility of a second date or the finality of it being a one time phenomenon with that date. A simple, casual text the day after would be apt for an evening you both might have enjoyed.

10 Tips for Approaching Women

February 11th, 2008 by nwoclone-army
Most guys get caught up in guessing what to say; here’s what they really should do
What do you do when you see a woman you are attracted to? Do you run and hide? Do you use some canned line that you read on the Internet? Do you stand there in fear trying to think of the right thing to say? What is the right thing to do?
When approaching a woman, most guys make the mistake of thinking too much about what to say. They believe there’s one magic line that will work in all situations. They rehearse this magic line, and when they deliver it, they hope the woman will become instantly attracted to them.
Unfortunately, rarely does this approach work — because most of what you say is irrelevant. To catch a woman’s attention, it is all about the confidence you display when approaching her.
Here are 10 surefire ways to intrigue her every time:
1. Observe something. Make a comment about something you observe in the environment. This is especially effective at the grocery store. For example, if she is ordering a turkey sandwich, ask her if the turkey is good here.
“Make your comment immediate to the situation”

Make your comment immediate to the situation and it will seem perfectly natural. No matter where you are, there is always something interesting to comment on.

2. Smile. This shows her that you are friendly and confident. A genuine smile not only feels good to you, but will put her at ease while creating openness in the interaction — a requirement for building rapport.
3. Do not hesitate. If you hesitate in your approach, this tells her that you are not feeling confident — an immediate turn-off. When you see her, walk over to her within a short period of time (the three-second rule). Show her you are a man who knows what he wants and goes after it.
4. Positive body language. If you approach hunched over with your head down, you are sending negative information about yourself, which makes you dead in the water before you begin. Stand up straight, with shoulders back and chest out, and use a firm yet relaxed walk.
5. Not too fast. If you walk over too fast, you could likely trigger her internal alarm. A calm, casual approach is usually the best way to make her feel at ease with you.
6. Keep eye contact. Never be the first to break eye contact when you approach. If you do, this sends the message that you are not feeling good about approaching. When you use strong eye contact, she will feel more drawn to you. With practice, you can master this.
7. Listen up. Make sure you pay careful attention to what she says. Do not have your response pre-thought out. Women love a man who pays attention to the details of what she says. If you start throwing out random words, she will lose interest fast.
8. Do not fidget. Fidgeting after you approach is distracting and shows you are uncomfortable. If you communicate that you are uncomfortable, she will feel uncomfortable, too, and will close up. Practice being aware of your movements. Pay attention to those movements, or lack of movements, that communicate comfort and confidence.
9. Lighten your tone of voice. The tone of your voice is a very powerful tool. Approaching her in a light and playful tone is one of the best ways to start. You could also begin in a serious tone, accusing her of something like "
“I hope you saved some turkey for me”

I hope you saved some turkey for me," followed by a quick smile to let her know you are joking. Practice playing with your vocal tone with your friends — notice the different reactions you get when you say the exact same thing using varied tones and fluctuations.

10. Lean away from her. A man who leans in too far when he talks often makes a woman feel crowded. A better approach is to lean away from her slightly. This lets her know that you respect her space, boundaries, and are comfortable with yourself.
The key to making these tips work for you is putting them into practice! Practice these tips and see the reaction you get. When you put them all together, you will be surprised at their power.

How to Meet More Women

February 11th, 2008 by nwoclone-army
Be the mayor of your own life
Whenever I do a dating boot camp, guys are always amazed that I seem to know everybody. The first time one attendee, Yakub, did a boot camp with me, he kept marveling at this. He started calling me "The Mayor of New York City."
It arises from the fact that everywhere I went, I knew somebody. I knew men. I knew women. I knew kids. I knew dogs. I knew shopkeepers. I knew everybody. By knowing all these people everywhere I went, I got to know more people.
“The number one mistake that men make when they go out and try to meet women…”

The number one mistake that men make when they go out and try to meet women… is that they go out and try to meet women. Think about that statement for a second.

You call your friend on a Saturday afternoon to go out and meet women. So what do you do? You go to a coffee shop, you stare at a good-looking woman, you try to remember the perfect opener… and you ignore everybody else in the place. Does this sound familiar? I used to do the same thing. It’s the reason why you do not meet women.

Become mayor of your own life

Now, let me teach you a technique that is far simpler and a thousand times more effective. It is not only more effective, but it will decrease your rejection rate by 100 percent.
Instead of going out and trying to "meet women," go out and start meeting people. That’s right.
“Start talking to everybody you know, and everybody you don’t know.”

Start talking to everybody you know, and everybody you don’t know. By going out and communicating with everyone, you’ll start attracting women instead of having to chase them.

By talking to the little old lady who is walking her dog, that hot woman you want to meet will notice the good positive energy you’re displaying toward the old lady. This will cause her to become attracted to you. She will stop, and she will most likely approach YOU.
Five people you need to meet
Here are five key people in your life you need to get to know:
  1. The checkout clerks who work at your local market
  2. The person who serves you coffee every day
  3. The person who works the counter at your dry cleaners
  4. The person who makes your sandwich at lunch
  5. The host at the local restaurant where you eat regularly
By getting to know these people, when someone you’re attracted to is behind you in line, you can break the ice by starting a conversation with the checkout person with whom you’re already comfortable. That eases the pressure of striking up a conversation with a new person.

No more routines

“You should be talking and friendly to everybody you meet.”

You should be talking and friendly to everybody you meet. By doing this, women are going to become attracted to you. Not only that, when you talk to everyone, you don’t look like one of those sleazy pickup artists who run lines and routines on women wherever they go.

I’m going to lose it if I hear one more guy walk up to a woman and say, "Can I ask you a question? Who lies more, men or women?" You know how I feel about routines.
You will never again need a routine if you naturally learn how to become mayor of your own life. Not only that, but women will start to approach YOU. So now, who would you rather be for the rest of your life — the pursuer or the pursued?

Obesity surgery seen as diabetes cure

January 23rd, 2008 by nwoclone-army

CHICAGO - A new study gives the strongest evidence yet that obesity surgery can cure diabetes.

Patients who had surgery to reduce the size of their stomachs were five times more likely to see their diabetes disappear over the next two years than were patients who had standard diabetes care, according to Australian researchers.

Most of the surgery patients were able to stop taking diabetes drugs and achieve normal blood tests.

"It’s the best therapy for diabetes that we have today, and it’s very low risk," said the study’s lead author, Dr. John Dixon of Monash University Medical School in Melbourne, Australia.

The patients had stomach band surgery, a procedure more common in Australia than in the United States, where gastric bypass surgery, or stomach stapling, predominates.

Gastric bypass is even more effective against diabetes, achieving remission in a matter of days or a month, said Dr. David Cummings, who wrote an accompanying editorial in the journal but was not involved in the study.

"We have traditionally considered diabetes to be a chronic, progressive disease," said Cummings of the University of Washington in Seattle. "But these operations really do represent a realistic hope for curing most patients."

Diabetes experts who read the study said surgery should be considered for some obese patients, but more research is needed to see how long results last and which patients benefit most. Surgery risks should be weighed against diabetes drug side effects and the long-term risks of diabetes itself, they said.

Experts generally agree that weight-loss surgery would never be appropriate for diabetics who are not obese, and current federal guidelines restrict the surgery to obese people.

The diabetes benefits of weight-loss surgery were known, but the Australian study in Wednesday’s Journal of the American Medical Association is the first of its kind to compare diabetes in patients randomly assigned to surgery or standard care. Scientists consider randomized studies to yield the highest-quality evidence.

The study involved 55 patients, so experts will be looking for results of larger experiments under way.

"Few studies really qualify as being a landmark study. This one is," said Dr. Philip Schauer, who was not involved in the Australian research but leads a Cleveland Clinic study that is recruiting 150 obese people with diabetes to compare two types of surgery and standard medical care.

"This opens an entirely new way of thinking about diabetes."

Obesity is a major risk factor for diabetes, and researchers are furiously pursuing reasons for the link as rates for both climb. What’s known is that excess fat can cause the body’s normal response to insulin to go haywire. Researchers are investigating insulin-regulating hormones released by fat and the role of fatty acids in the blood.

In the Australian study, all the patients were obese and had been diagnosed with type 2 diabetes during the past two years. Their average age was 47. Half the patients underwent a type of surgery called laparoscopic gastric banding, where an adjustable silicone cuff is installed around the upper stomach, limiting how much a person can eat.

Both groups lost weight over two years; the surgery patients lost 46 pounds on average, while the standard-care patients lost an average of 3 pounds.

Blood tests showed diabetes remission in 22 of the 29 surgery patients after two years. In the standard-care group, only four of the 26 patients achieved that goal. The patients who lost the most weight were the most likely to eliminate their diabetes.

Both patient groups learned about low-fat, high-fiber diets and were encouraged to exercise. Both groups could meet with a health professional every six weeks for two years.

The death rate for stomach band surgery, which can cost $17,000 to $20,000, is about 1 in 1,000. There were only minor complications in the study. Stomach stapling has a 2 percent death rate and costs $20,000 to $30,000.

In the United States, surgeons perform more than 100,000 obesity surgeries each year.

The American Diabetes Association is interested in the findings. The group revises its recommendations each fall, taking new research into account.

"There is a growing body of evidence that bariatric surgery is an effective tool for managing diabetes," said Dr. John Buse of the University of North Carolina School of Medicine in Chapel Hill, the association’s president for medicine and science.

"It’s just a question of how effective is it, for what spectrum of patients, over what period of time and at what cost? Not all those questions have been answered yet."

Medical devices used in the study were provided by the manufacturers, but the companies had no say over the study’s design or its findings, Dixon said.

___

Is She Attracted to Me?

November 20th, 2007 by nwoclone-army

Is She Attracted to Me?

All the signs are there — if you know how to read them

By David Wygant Sepcial to Yahoo! Personals

Updated: Nov 20, 2007

 
I get so many emails from men asking me this question: "How do I know if she is attracted to me on date?"
This a great question, and knowing the answer will make going in for that kiss become her idea and not yours. That’s right, guys — the first kiss has to be her idea. She has to want it and desire it, and learning what her body language is saying is key to the first kiss and knowing if she is attracted to you.
When you are out on a date, sit across from her at a table, or if you are about to sit in a booth, let her sit down first and see where she invites you to sit. Some women will invite you to sit right down next to them in the booth and some will not. If she invites you to sit down next to her, she is telling you that she has an initial attraction to you.

Be a listener, not a talker

“The first step to making the date a true success is to listen to what she is saying.”

The first step to making the date a true success is to listen to what she is saying. Pay attention to the details, and react to what she has to say. That does not mean that you can’t share a story or two about yourself, but the best dates are the ones where you’re in a listening and reacting mode rather than a talking and bragging mode.

Of course, she wants to hear about who you are and what you are all about. She does not want to hear you brag about how much money you have or how successful you are. She also does not want to know about your negative dating history.
She wants to know about what you have learned in life to this point.
“She wants to see how positive a person you are, and she wants to imagine being able to hang out with you.”

She wants to see how positive a person you are, and she wants to imagine being able to hang out with you.

There is a lot to learn about a first date. I really suggest you book one hour of phone time with me to go over all of this. So if you desire to be the guy who women want, then email me right now and let’s book that hour.

How to read body language cues

Now what type of body language should you be looking for when you’re sitting across from her at that table?
Pay attention to her eyes. One sign of attraction is when her eyes are open really wide, and her pupils are enlarged when you are talking or when she is talking to you.
Another sign of attraction is that when you are speaking, she will lean her body into you and literally be drawn in with your words. She will not get up and go to the bathroom. She will sit there and not want to miss a single second of the date:
  • She will not look at her watch.
  • She will not look around the room.
  • She will be totally fixated on you the whole time.
  • She will play with her hair and lick her lips before she moves towards you, because she is creating a sexual feeling inside her.
  • She will reach her hand across the table and glance at yours.
  • She will touch your shoulder or another part of you very casually.
  • When she is speaking, she will touch her leg or her face, imagining it was you who was touching her.
There are many others, but these are a few good ones to get you started.
“Keep in mind that what she is doing and communicating with her body is on a subconscious level.”

Keep in mind that what she is doing and communicating with her body is on a subconscious level. She is not aware of what she is doing, and that is what makes this so powerful.

One last thing: How do you know after all of this that she wants the kiss?
She wants the kiss if, when you walk her to the car, she lingers and keeps talking and looking at you. What you do then is go in, move towards her lips, and see what she does. Then pull to the side and give her a hug. This will create tension. If she then talks more, look at her, touch her face and move in for the kiss.