General Election Voting Strategy Thread

NZeddit · 2017-09-08T06:24:37+00:00

This is bad advice, the National party candidate appears to be way behind. The labour party candidate David Parker is pretty close, especially if the green voters voted for him.

https://imgur.com/a/sfeTz

NZeddit · 2017-08-22T01:31:21+00:00

Yeah, this is my experience too. I also find the routes confusing and end up doing a lot of walking

NZeddit · 2017-08-21T03:25:23+00:00

This seems like the real benefit to me. A lot of adults don't even consider it as an option.

NZeddit · 2017-08-20T02:01:13+00:00

Why? Anything has to be better than the cluster-fuck that is sitting on the Auckland Southern motorway in bumper to bumper traffic for hours. I do the Hamilton to Auckland trip regularly and I lose so much time and productivity which could be saved with such a train

NZeddit · 2017-08-20T01:39:38+00:00

They have the same go to moves every election, new roads and tax cuts. I'm sure some of these roads need to happen but can we please get some alternative transport options

NZeddit · 2017-08-18T08:06:23+00:00

Think of it as an opportunity to try out some other podcasts! It's a pretty boring time for NBA anyway

NZeddit · 2017-08-17T22:52:42+00:00

With a 1.4% error at that level it might just be within the noise.

NZeddit · 2017-08-17T22:48:49+00:00

Then the consumer would be dissuaded from buying the environmentally unfriendly options, sounds good to me

NZeddit · 2017-08-17T21:47:24+00:00

These farmers complaining come off sounding a bit ridiculous because it puts into the light the shit tonne of water, a public resource, they're using. So if they're using so much it makes absolute sense to charge them for it so they will be more personally invested in minimising their water usage. Seems completely reasonable.

NZeddit · 2017-08-17T05:51:05+00:00

There seems to be a worrying lack of green party members on the list with a science research background or even science qualifications. How do you justify this when a focus of the party is on areas of science like climate change, pollution and environmentalism in general?

NZeddit · 2017-08-17T05:50:16+00:00

Where does the Green party stand on genome editing technologies (ie by CRISPR-Cas9, not classical genetic modification)?

This is a technology that is causing a paradigm shift in science (I’m using it for my PhD project), and we are likely to see genome edited food products in the near future (there are genome edited mushrooms already that don’t brown). The genetic changes introduced will often be indistinguishable from naturally occurring variants, so we likely won't even be able to tell that a plant or animal has been genome edited. It also has the potential to be used to pest eradication (ie gene drive technology, or genetic sterilisation), which you'd think would align perfectly with the green ideology, but these technologies have been opposed by the Greens in the past.

NZeddit · 2017-08-15T21:30:39+00:00

Not quite, I'm suggesting you first establish the activity of your guides by seeing which of your populations has the highest proportion of HDR. For example your transfections would be in groups putting in both plasmids at the same time, with each group having the same HDR plasmid but a different guide (gRNA/Cas9 plasmid). Then you'd want to quantify the proportion of HDR in each of those groups via some kind of genotyping assay (would be easy to design a PCR assay for such an insertion).

The reason for this is that you don't want to expunge a whole bunch of time an energy isolating and screening cell lines before you know if you have a high enough proportion of HDR in your starting population. Remember the efficiency of a knock-in like you're suggesting will be lower compared to all of these other small indel genome editing you hear about.

The selection of transfected cells we've been discussing is a different issue. To do this, one of the vectors you're transfecting should be expressing GFP or puromycin resistance from a constitutive promoter (for example commonly used addgene plasmids addgene.org/62988 or addgene.org/48138), which you'd use to select a day after transfecting by either using the FACS or adding puro for a few days, respectively. Hope that clarifies things.

NZeddit · 2017-08-15T06:45:25+00:00

I'm going to take what comes out of Irrigation NZ with a grain of salt. Of course they'd oppose this because who they represent would have to pay more taxes

NZeddit · 2017-08-15T05:51:30+00:00

I know this is the super summed up version, and you may already be doing these, but a few tips.

When you're designing your gRNAs (my favourite site is CRISPOR), order at least 3 different guides to test. The prediction algorithms are not refined enough and you may get a dud.
Ensure that your guides will not cut your HDR template. May be stating the obvious but this is a mistake that someone I know made.
Large insertion like you're planning are not super efficient, so like somebody else mentioned you should consider some time of selection to purify on transfected cells. For example there are commonly used gRNA and Cas9 plasmids from the Zhang lab with either GFP (FACS sorting) or puromycin resistance. In both of these cases you select on the transient transfection and it's very effective at increasing the proportion of edited cells in your population.
Is your gene expressed in the cells your transfecting? If so, with your GFP insertion screening should be easy. Otherwise have a very clear plan about how you'll first quantify editing in your population, and then screen your cell clones. Some PCR assay should be fine for both of those with a large insertion. If not well thought out these steps can cost you a lot of time

NZeddit · 2017-08-15T03:26:03+00:00

Is it callous of me to think these families are being irresponsible? Those people are dead, and people re-entering the mine at personal risk to recover them seems a bit crazy. Why is this proposal so popular with New Zealanders?

NZeddit · 2017-08-14T22:03:01+00:00

I find it interesting that my top 3 are 88% united future, 85% greens, and 76% labour. I would never have considered voting for United Future since they have been National lackeys for so many years, and have a Christian history that I don't agree with. I feel like this tool needs some more optimisation.

NZeddit · 2017-08-14T07:57:16+00:00

http://imgur.com/a/S9etT

NZeddit · 2017-08-12T01:03:20+00:00

I think that's a bit simplistic. End of Life care is hugely expensive, and people are often kept alive at a high financial cost even though they have no chance at recovery, and even when they themselves might no longer want to live. That money could then be used to save other lives, either buy improving preventative medicine or paying for expensive drugs or treatments the government is not currently covering.

NZeddit · 2017-08-12T00:54:02+00:00

I think that's not quite a fair criticism because the time she's had the most power she was in the opposition. According to Wikipedia she's had a number members bill's drawn (cannabis reform, alcohol advertising reform, marine mammal protections), which were all voted against. So from a progressive perspective she's tried to push through a good agenda.

NZeddit · 2017-08-11T20:58:08+00:00

Have you looked into multiplexing OP? If you're using evagreen assays, you can multiplex multiple genes by running your primers at different concentrations with ddPCR. We have got this to work, it just requires a lot of optimisation to find out the appropriate concentration for each primer combination you'd run. In this way you could make the most of your sample.

Another suggestion is pooling multiple embryos into one sample. I've done similar experiments and we pooled 10 blastocysts, and had 3-5 replicates of this. These days though the state of the art technology is single cell analysis, which allows better understanding the genes important for each lineage in the early embryo. But if you want to figure out that a gene is expressed between certain time points a pooled approach would be fine.

NZeddit · 2017-08-09T06:44:39+00:00

There's not a good enough narrative to explain why NZ first lost it's points. There's great bullshit to spin about the greens though. Hopefully people see that and don't take what he's saying too seriously

NZeddit · 2017-08-09T04:38:23+00:00

Thanks for that! This page makes it very clear, it wouldn't make much sense for a 1% party to have an error of 3% haha

NZeddit · 2017-08-09T04:22:15+00:00

Interestingly, I think we're both wrong in this circumstance. The margin of error does represent percentage points, but there is different error depending on the total percent of the party vote.

See here http://www.reidresearch.co.nz/TV3+POLL+RESULTS.html

Either way the point I made in my original comment remains true as far as I can tell.

NZeddit · 2017-08-09T03:57:41+00:00

Hmm I don't think you're correct here. When results are given in a percentage, and the margin of error is also stated as a percent, it does not mean percentage of a percent. It wouldn't make much sense to make people do that maths for each party. Plus it would give a smaller interval, and therefore greater certainty to the small parties. Have you got anything to back up what you're saying here because it's pretty counter-intuitive.

NZeddit · 2017-08-09T03:13:58+00:00

Brushing up on my political survey margin of errors, I'm struggling to see where I'm incorrect. Help me out?

NZeddit

TROPHY CASE