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Looking for female and male nepenthes leaves for sexing experiment

I am soliciting for leaves from known male or female Nepenthes. I promised someone female plants from my Nepenthes sprouts and I intend to keep it. This is a PCR based assay from a 2007 published paper. The techniques utilized are commonplace but the DNA was never sequenced by the researchers. I intend to replicate the protocol, clone, and then sequence the "male specific band" if there is one. I am looking for a small subset of leaves.

Stipulations
1) You must be able to validate the sex of the plant (i.e. have seen it flower. Clones from a TC "male" is not good enough)
2) For every verified leaf, you can send me (1) unknown of your choice which will be in the experimental group

I would prefer growers who live in the bay area. However, I am open to all, as genomic DNA is very stable.

Jen
 
i see a lucrative opportunity here... best of luck on your efforts Jen.
you might want to go to the SF Conservatory of flowers to get a few samples if they allow it, multiple plants there are flowering, so you can verify the sex of the plant. sorry i have no samples for you to work with...
 
have known female ventricosa, maxiama. Known male ventricosa. known female ventricosa hybrids. etc
 
This is just for fun, so to speak, as I am done with making my own PPM. This is next thing on my list of things to do. This will be done in parallel with some trnk species ID. Andreas seems to have left out Edwardsiana in his paper, so I will have to do that as well. I can always make the drive and ask Peter for some leaves from his personal collection. At least there is his cat to play with there...

Thanks for the offer Kulamauiman. I am guessing you also have some wildcard plants that you would like to know the sex of?
 
This is just for fun, so to speak, as I am done with making my own PPM. This is next thing on my list of things to do. This will be done in parallel with some trnk species ID. Andreas seems to have left out Edwardsiana in his paper, so I will have to do that as well. I can always make the drive and ask Peter for some leaves from his personal collection. At least there is his cat to play with there...

Thanks for the offer Kulamauiman. I am guessing you also have some wildcard plants that you would like to know the sex of?

dunno. most of the mature plants I bought came with known genders. Two haven't flowered but, I think I need to let them vine more for that to happen. Do ahve some oddballs but. eh. neps are not really my thing. I just grow to annoy people by saying the all look the same to me :p
 
i gots a couple males. pm me
 
I have a female N.smilesii and a male N.spectabilis for you to test, and am gonna toss in a seedgrown hamata leaf and ovata leaf for sexing. PM me with your address and stuff when you have the chance. :)
 
Don't tell me that is one of BigBella's hamatas - that would be so ironic. I will send you the details tomorrow.
 
I actually have one of his hamatas :p but it can wait. sort of like Christmas presents.
 
  • #10
I have a verified female ventricosa. That's the only nep of mine that I know. PM me if you want samples.
I've thought of doing something like this before too; if only I had the lab experience and resources. Exciting to see someone doing it!
 
  • #11
I've got a splendiana female and male red leopard. Then of course a slew of unknowns. :p
 
  • #12
I have verified male aristolochioides, male hamata, male predator, male vent x lowii.. in short, lots of males. I would be interested in finding out the sex of my edwardsiana.

Edit: missed this "1) You must be able to validate the sex of the plant (i.e. have seen it flower. Clones from a TC "male" is not good enough)" while the plants I have mentioned havent flowered for me yet, they have all been labeled as male by the person who took the cuttings. For the hamata, all AW clones are male, the aristo clone has flowered as a male for pablo, the predator has flowered for the person who I got the cutting from, and the vent x lowii was labeled by leilani as a male plant. I hope I can still be of some help for this.
 
  • #13
I have quite a few females that have flowered for me that I'd love to give tissue for this experiment! I have a female N. Spathulata, N. thorelii X truncata X campunalata, N. Hirsuta X Tiveyi, and a female N. campotiana X ?. I also have a male N. X Aeneas which I could give tissue for. I've got a few vials I can put the tissue in with some water to keep them fresh... would this work for you? I'd love to help out with this. I only live about 5 hours south of the Bay Area so the cuts should be pretty fresh on arrival.
 
  • #14
Hello,
Initially I was looking for n=10 in the experimental group with 5 males and 5 females. The 5 controls for each sex is not necessary, just for redundancy. If those interested could list what they would like to donate as exampled below:

Female
1. Pokie - N.Spectabilis

Male
1. Pokie - N. Inermis

Unknown
1. Pokie - etc..

Samples requirements
1) A small leaf is all I require. Usually for genomic DNA isolation 100mg or less is required but I would rather have enough sample for a couple iterations than ask for another leaf down the line. This is impart due to the high phenolic compounds present in Nepenthes and I may have to do further purification before the DNA is clean enough for PCR.
2) Young leaf - DO NOT GIVE ME THE OLDEST LEAF! Fibrous tissue will make me hate you, in essence. I would like to be able to do this with a dounce homogenize or motor and pestle + liquid nitrogen. I don't want to have to break out the big guns.
3) Please package the leaf in some fashion so that it will be alive as possible when it arrives on my doorstep.

When the list is full, I will send everyone PMs.

Heli, I can toss your eddie into the experimental group. It would be nice to have more than one sample to perform the trnK sequenceing for eddie species ID.
 
  • #15
Female
1. Exo - N. smilesii

Male
1. Exo - N. spectabilis

Unknown
1. Exo - N.hamata, ovata
 
  • #16
best means for sending the leaves would be in a folded paper towel moistened with RO-H2O, placed in a ziplock bag and that being placed in a small bubble package.

remember to properly label your specimens!
 
  • #17
Female
1. Exo - N. smilesii

Male
1. Exo - N. spectabilis
2.Pebes-N. aristolochioides
3.Pebes-N. talangensis


Unknown
1. Exo - N.hamata, ovata
2. pebes- N. edwardsiana AW, N. singalana



thanks
 
  • #18
Female
1. Exo - N. smilesii

Male
1. Exo - N. spectabilis
2.Pebes-N. aristolochioides
3.Pebes-N. talangensis
4.Heli- N. predator

Unknown
1. Exo - N.hamata, ovata
2. pebes- N. edwardsiana AW, N. singalana
3. Heli- Burbidgeae sg, edwardsiana sg

Do we send you a whole leaf or can we send you a piece of a leaf?
 
  • #19
Female
1. Exo - N. smilesii
2. lance - N. Hirsuta X Tiveyi
3. lance - N. thorelii X (truncata X campunalata)

Male
1. Exo - N. spectabilis
2.Pebes-N. aristolochioides
3.Pebes-N. talangensis
4.Heli- N. predator
5. lance- N. X Aeneas

Unknown
1. Exo - N.hamata, ovata
2. pebes- N. edwardsiana AW, N. singalana
3. Heli- Burbidgeae sg, edwardsiana sg
 
  • #20
Amusing. 2 eddies on the unknown. The impetus for this is actually to determine some UHL sprouts, Edwardsiana being one of them.

Lance: no unknowns for you?

Heli: For your own edification and so that you can make a volitional decision - PCR is a method of amplifying a specific DNA sequence by use of a thermostable DNA polymerase and short DNA primers that flank each side of the target sequence. For each cycle, the theoretical amplification is 2^N, where N is the number of cycles. When amplifying from a small amount of DNA, I usually perform 30-40 cycles as a starting point. Theoretically, there are methods of amplifying from less than one molecule. However, I like to exist in the real world and not imaginary. If there is no signal, I usually increase the number of cycles. Now, the more cycles, the more one invites DNA base pair errors. The DNA polymerase has an inherent error rate and the one that is used for amplification of dirty DNA, Taq polymerase, is the most error prone. There are often false positives and false negatives in PCR when performing higher number of cycles. The less starting DNA in the tube = the more cycles will have to be done.

Now, extrapolating this to real world applications, rapist and serial killers are some times convicted by DNA fingerprinting with PCRs that have undergone 90 cycles. I can vouch that some times I see Jesus in my gels after 90 cycles. Of course, this is only an overly simplistic view of PCR. Everything in life is so much more complicated...
 
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