dumbdumb

just kind of curious.

Forgive the brevity of this story: I married a forgein girl I loved in college. We never consummated our marriage sexually. She was pregnant by a guy before our marriage. The baby is NOT mine. I was mad and left her, but we never got divorced. I was a sucker for her hardship stories and I agreed to apply for a green card, for her benifit, and she got one. After that I heard little from her for 4 years. We have recently have been in contact with eachother via phone. She is now eligibe for USA citizenship, and is asking for my personal information (all the stuff she lost in Katrina).

She has a green card. I'm reluctant to give her my personal info. I am hoping she can apply with no further obligation on my behalf. I want nothing to do with her or her child, and have started my own new life, but can't afford to divorce her. Question: If she obtains citizenship, will I be obligated to her or her child (who does not have my last name), finacially? I CT, she TX

Advice Please!

Hi everybody,

I bought Hyssop "Jubilee" last year for some friend's garden last year. We loved its perky green leaves, and its brilliant tall flowers. I left it to seed over winter, which it did splendedly.

I try to visit their garden once a week, and all seemed to be going well.

Today, I dropped off some castor bean plants, and took a two minute tour of the garden.

I noticed the Hyssop had a lot of brown spots on the leaves. I took a further look, and noticed some of the leaf skeleton showing. Upon turning the leaf over, I saw some baby bugs munching away (actually they dropped immediately to the ground).

I suspect it's what I call "earwigs".

Any ideas on how to proceed in correcting this early devastation? Or should I just leave em be? HELP!

Thanks for your thoughts and ideas.

Ok, Ok.

It's Friday night and I just did a DNA extraction in my own kitchen.

I'm a true NERD!

I dissolved some sea salt to about saturation in a glass.

I dry brushed my teeth and tongue and cheeks.

I took a small mouthful of the salt water, and spit it into another glass.

I mixed two squirts of soap and some water from the faucet.

I did a 50% mix of soap water with the cell/salt mix. I swirled for a couple of minutes. I added 3 times alcohol to the soap/salt mix and gently swirled.

I see what I think is DNA (looks like a big wad of phlegm).

Is that DNA?

I'm used to commercial kits where cell debris is spun out as a pellet.

Did I do this right?

Thanks for laughing with me!

Sometimes I find myself just following protocol, without really knowing why.

Is there any reason for using 0.5X running buffer, when my gel is composed of 1X buffer? (I mostly use TAE buffer, and when called for I also use TBE buffer).

I am guessing its just to save a nickel on buffer costs, but maybe there is a REALLY good reason.

And, since I have your attention, I'm wondering if any one knows of a good size marker that is good at differentiating PCR products sized between 1kb and 1.5kb. Mine is just okay, but would like something really more definitive in this range. Any comments, thoughts, rude noises....all are welcome!

Best to you!

I'm craving Macleaya Cordata!!!!

I know it's pretty invasive, as I used to grow it in some pretty tough spots, but I have a new (and even tougher) spot in mind.

But I will not be able to dig up my original plant.

I am looking for roots and a bit of leaf from gardeners who know they have plume poppy and are going to dig it up to make space for better plants.

Here is a brief summation of the plant, just in case your not sure if you have it growing in your garden:

Macleaya cordata "Plume Poppy".... A true drama queen. 6' - 8' tall plumes of airy white flowers in July and August over blue-green scalloped foliage. Self-sows like crazy! Adds an instant eye-catching mass to any border.

Looking to own 3 or 5 roots with fresh greens but will accept and trade what ever comes my way.

The season is early but I know I am flush with some good garden stock from last season worth trade.

email me!!!

Thanks for sharing!!

I've been answering calls from 1-800 numbers, and the person on the other line hangs up after I say hello. Its starting to freak me out, as this has been going on for some time now.

I called the number back but no response. I'm on the national "no call" list.

Somebody is calling me for non-business reasons and I would like to know who it is.

Is there way to find out for free who the number is connected to?

If one takes genomic DNA, cuts it with a "four base" restriction endonuclease, is cleaned up, labeled, and then hybridized to a micro array chip with probes (say 50 bp long on average) representing the whole genome, is there a limit to the size of a fragment a probe can hold on to?

For example, can a 50bp probe "hang onto" a 50kb fragment, even when the 5' to 3' sequence of the fragment matches perfectly to the 3' to 5' sequence of the probe?

What happens if the probe corresponds to the sequence right in the middle of a 50kb fragment? Will the probe hybridize to the middle of the fragment ? Would there be any impact on measuring the fluorescent label?

Can anyone suggest papers dealing with the dynamics of fragment/probe interactions?

Thanks for you help!

I have been out sourcing my sequencing from cloned DNA. My inserts are about 1kb to 1.5 kb. My inserts are "cleaned up" PCR amplicons generated from bisulfite treated genomic tumor DNA.

I ask the company to pick 8-12 colonies, grow them out, purify the insert, and sequence using forward and reverse primers (I do this because my inserts are bigger than their one way read outs provide).

They give me okay reads with the reverse primer (like 5 of 8 colonies picked will give a 60% alignment at best). But I'm sort of new to this company, so I don't know what to expect.

But the forward primer will mostly result in "NNNNNN" reads.

After a little research I found out that bisulfite treated DNA may result in "hairpin" structures, which makes sequencing more difficult. But from one clone, I usually get one good read with the forward primer.

Why can't I get readouts from the other 7 forward primer sequences?

Any Bisulfite Sequencing Experts?

I'm using TA cloning kit from invitrogen. The plasmid is pCR (if memory serves me properly).

I amplify the amplicon from bisulfite treated DNA, and cut my band out of LM agarose. I perform a gel purification, and perform an overnight ligation. I do a transformation the following day, and grow the bacteria out on an ampicillin plate. I see mostly blue colonies, and some white colonies.

I send out the plates (overnight delivery) to a Texas company: Seqwright. They pick white colonies (8-12) which I have circled for them and grow out in LB media containing ampicillin (maybe kanamycin, too, I not sure).

On one occasion Seqwright has been unable to grow out the picked colonies. All "inserts" were batched in the way I described above. All failed. Other batched inserts processed in the same way, are able to grow.

What did I do to that one batch I sent out for sequencing? Why did white colonies not grow out in media. (I triple checked all steps).

All advice is welcomed.

Hi everybody. Thanks for being here for me.

I could afford a mini-christmas tree this year. And I LOVE it. it's a 2 foot rosemary shrub. It smells better than the pines I remember vividly as a kid.

However, mine seems to be browning out, with its needles turning black.

I water thoroughly and allow to drain thoroughly, and set it up in the sunniest window (where ever other plant seems {okay for now}). I water daily, as the directions suggest.

My simple device for measuring indoor temperature reads 80 degrees.

I do my very best not water the needles.

Some water landed on the lowest of boughs, but the black needles extends all the way up.

She is not dead, just loosing needles faster and faster since I adopted her.

Any suggestions as to how to help her thrive?

One more question, Is it OK to recycle her dead needles among my other plants? Sort of like an indoor mulch?

Thanks for your help!

(I have more questions on the way, but will save for another day)

I have bad psoriasis.

I've read papers on helping people with psoriasis lately.

Interestingly it has to do with STAT3.

Simply put, STAT3 is a signal transduction system in cells with an output of transcription factors that are involved in maintaining psoriasis.

I read that decoy oligonucleotide therapy helps with this.

Here are my questions.

If I purchased two oligos and encouraged them to anneal, and then applied them directly to my skin (in either water or EDTA), would the DNA enter the cell topically? My oligo sequence would be for STAT3 transcription factor.

Do I need a sulfur group to inhibit DNA Nuclease?

Is there another factor for cell-mediated incorporation of DNA into the cell cytoplasm, and then the nucleus, that you know about?

What other things should be considered with topical DNA oligo's?

Mine is pretty painful. I have it on one elbow, and as a kid I had it all over.

I still don't fully understand the physiology of why it comes and goes.

HELP!

I am helping out a friend buy a car. He has the Nissan Versa Hatch SL CVT in mind now.

He asks me a million questions. Some I don't know the answer to. He does way more research than I do. I want him to buy a car, but sometimes I end up giving "any answer", and that sits on my conscious.

I will ask some of his questions here. I want to ask his concerns about extended warranties. Does he really need one, etc.

He claims if you buy one (for a new car), you should NOT buy from the dealer. Because, in the world of finance, the underwriter of the policy could close shop before your policy ends. And legally you have no right to claim.

I recommend Him to always buy from the dealer (I never had a problem)

I questioned where he gets this info from. Let me give you this link: http://www.carbuyingtips.com/

I checked some of it out. I don't know. Its kinda interesting BUT it also looks like internet info-mercial.

Any alternative sites? Any comments on insurance (also GAP)

I think I just want to tell my story. It is not so sad, but I am sad because of it. here is a bit.

I moved to a new state, knowing no-one. I fell for a guy, we had an on-and-off relationship for 12 years.

I broke up with him 9 months ago.

I thought I would be over the depression by now.

Hello, its new years eve. I don't mind being alone so much, but it kills me that im still thinking about him. not just today, but everyday after 9 months!

my main goal for dropping him: he consistently showed preferential treatment with his friends over the years. this made me not like his friends. which made them not like me. I should mention: his popularity always superseded mine in the bar scene.

I thought he was my soul mate, because, despite the preferential treatment, he loved me physically and mentally as I never experienced before.

I am home alone tonight and every night since our break up. I have a very small group of fair weather friends left.

I'm sad. talk me out of this. HELP

Just wondering: if my fuzzy legs touch the pollen on my blooming Amaryllis, and then they rubbed some of the pollen onto the stamen, would I be enhancing the production of a new bulb?

How do new bulbs form anyhow?

Thanks in advance for you responses!

My best friend has a beat up car that needs to be replaced!

I've have worked HERO hours to find him a car within his specifications: hatchback, low mpg (he says hybrid, but he liked a few non-hybrid cars), and gently used. He thinks his purchase will support anti-Alaskan oil devoplement.

I take his points in stride. BUT THE KID NEEDS A NEW CAR!!

I took him to many lots, and he always walks away. I know his reasons, and I understand them too. He doesn't want to get burned in bying a used car.

He thinks hybrids are tax free our state(CT)? and found one at a "c_r_ax" dealer" [the other two letters are a and m] which i leave out for more honest opinions.

I keep warning him not to buy used, especially from "c_r m a x" !!!!, because they have a bad reputation.

I guess I'm hoping to prove him wrong, but in fairness I am looking for impartial advice either way (and i'm not really looking for c_r_max sales people's bias on the subject, other opinion's welcomed).

more later.

Any technical help for cropping holiday pics that will fit into 2.5 x 3.5 frame size? I am working with iphoto on a mac, but willing to download other working applications that will give the size i need.

I take pcr amplicons and put them in a vector, then put the vector into e.coli. I grow the e.coli out on a plate then pick a colony to transfer to liquid agar.

I thought this was cloning, but then I ran across this term "sub-cloning" in a job description. Whats the difference?

It recently occurred to me that I missed out on understanding this phenomena called chimerism.

I recall seeing a few specials back in August 2006 about Lydia Fairchild on ABC networks.

I didn't quite understand the big "theme" here, to explain how the kids share no genetic identity with their biological mother.

I have followed up recently on the story, reviewed the story, and it is still not making sense to me.

Does anyone have a website to share, which might show the pedigree for this phenomena?

Or, would anyone be willing to explain to me how Lydia Fairchild's haploid genome was swapped with someone else's genome?

The concept of "you are your own twin" would be made clear after testing both parents of Lydia, right? Or did I miss in the story that one parent (or both) were not available for genetic testing.

Thanks for your help!

Here is a link, in case you forgot the details regarding this story.

http://abcnews.go.com/Primetime/story?id=2315693&p...

I'm definately no expert here, but very curious, as it applies to my current job.

Can they incorportate where ever they want to in the genome?

Or are their "sequence" rules that viruses follow?

Or is it species dependent?

Thanks for your input.