Talk:Northwestern blot

Review by Neelix

It looks like you've added some good information to this article. I have some recommendations about how to build upon what you've started:

1. It would be a good idea to add paragraphical content to the empty sections.
2. Categories are very important for Wikipedia articles; you should add as many as are relevant.
3. The capitalization in the title and the article's body should be consistent; use whatever capitalization is most common in the literature.
4. There should be no unsourced content in the article; the information in the "Origin" section should be sourced or removed.
5. Is the Molecular probes template relevant to this article? If so, a link to this article should be added to the template and the template should be added to the article.
6. Images would make this article more visually appealing. Perhaps one of Edwin Southern?
7. A "Further reading" section listing books on this subject would be a good addition.

I hope you find these suggestions useful. Let me know if you have any questions. Neelix (talk) 04:20, 31 March 2014 (UTC)

Hi Neelix, Thank you for the review and suggestions. All good points, and we'll look to address them. Regarding the paragraphical content, you're aware of the deletions that occurred. I was actually able to undo the last few revisions to get the content back. I then remade the subsequent revisions that I undid. So, I think we are ok on that front again. Thanks again.Msmrugby (talk) 11:24, 2 April 2014 (UTC)

Neelix and Msmrugby, I'm working on redoing some of the wording throughout the article to make it flow better. I've also added a section where reference protocols can be found. I'm working on trying to find some pictures of blots or the steps in the procedure, but I m finding it a little difficult but I'm going to continue looking. Tmckenne (talk) 01:41, 7 April 2014 (UTC)

Comments from Rmiller587

Great job starting a new article! It seems as though you've got a good layout to start with. Do you plan on adding more sections? A section about the type of data generated and how to analyze it could be useful. I know you've just started writing it, so it's probably not useful for me to tell you that you should add more information. I'd like to see pictures of how a Northwestern blot is setup if you can find any. Links to available protocols would be useful too, perhaps from a website like bio-protocol.org. Your first reference also seems to have a good protocol. Good job with wikilinking and the see also section! Rmiller587 (talk) 18:41, 31 March 2014 (UTC)

Thank you for the review Rmiller587. We had content from two of our sections that was deleted. I have since been able to get it back, but you did not see it when you reviewed unfortunately, so our article looked incomplete. Good suggestion about the additional section and addition of images. I have a few ideas for images that I'll be adding. Thanks again for the review and we'll address these points.Msmrugby (talk) 11:28, 2 April 2014 (UTC)

You're right, I thought you guys just got busy and didn't have time to work on it. The article looks much better now. Here are some comments on the parts I missed earlier...

You start off by mentioning RNA binding proteins but where do those come from and how does a researcher isolate them? From what are they isolated?

There are other things you can use instead of nitrocellulose, PVDF for example.

I think you should describe the importance and function of the blocking step.

I suspect the competitor RNA is artificially tagged with some sort of probe, fluorescent or radioactive (probably fluorescent now) but it's not mentioned.

What's the blot washed in?

The first two sentences in the Applications paragraph say the same thing and I think the writing in that paragraph overall needs work. Rmiller587 (talk) 13:52, 3 April 2014 (UTC)

Thanks for the additional comments Rmiller587. We will be addressing your additional comments in the Applications Section and the Techniques section. We're going too try and find relevant images that we can include showing the actual blot and/or the protocol if we can find one. I've since added an image of Edwin Southern as a starting point. Msmrugby (talk) 21:33, 6 April 2014 (UTC)

Maybe there are some images on the polyacrylamide gel electrophoresis article you can borrow? Rmiller587 (talk) 03:11, 16 April 2014 (UTC)

Great job with your continued additions! There's a lot of useful information provided and the layout is very easy to navigate. You've done a great job writing at a level that is understandable, even for those who are not molecular biologists. Rmiller587 (talk) 17:51, 28 April 2014 (UTC)

Thanks for checking back Rmiller587. Much appreciated!Msmrugby (talk) 03:56, 30 April 2014 (UTC)

Comment from Alpha centauri b (talk) 17:39, 2 April 2014 (UTC)

this article looks much better now with several separate sections. It would be great to see some figures or pictures of the techniques. Alpha centauri b (talk) 17:39, 2 April 2014 (UTC)

Hi, I had a few more ideas about the article.

• An intro subheading would be helpful.
• The "Origin" heading, this seems more like a description of the naming origin than an origin of the technique itself. Maybe renaming it something like "naming convention".
• I think an origin section would also be helpful. Who first originated it and when?
• In the application section I think you meant to say, "the blot could =be= analyzed..."
• Also one sentence might sound better as, "the location and concentration of the RNA binding protein on the blot will affect the result". What do you think?
• Some examples of the technique being used.

Alpha centauri b (talk) 18:48, 4 April 2014 (UTC)

Thank you for the feedback Alpha centauri b. Regarding the intro subheading, after looking at other wikipages, it did not seem like an intro section was the norm, so I decided to leave that out. Regarding the "Origin" section, the Northwestern blot technique ultimately resulted from the Southern blot, so in my opinion, it is an origin. To your point though, I am trying to find out if there is one individual that is credited with developing the Northwestern blot and will include this information if I can find it. We will be addressing your other points. Thanks for the feedback!Msmrugby (talk) 21:33, 6 April 2014 (UTC)

Alpha centauri b, thank you for the input on the article. It is really good to get other people's input on the article. I agree with you that rewording the sentence as you suggested will help with the flow more, so I changed it accordingly. I'm also looking for more examples of how the technique is being used. Tmckenne (talk) 00:47, 8 April 2014 (UTC)

I like that you added the image of Southern. Alpha centauri b (talk) 01:46, 8 April 2014 (UTC)

Thanks for checking back Alpha centauri b!Msmrugby (talk) 03:33, 16 April 2014 (UTC)

Review by martinhyou

Great job so far! Here’s some comments based on what I’ve read:

Introduction

• I really like the introduction. It’s easy to understand and to the point.

Origin

• Are you allowed to link to an article within the article and tell people to go to it to see an image? If that’s the case, it might be easier to find a free image somewhere and put it directly on the wiki page. Otherwise, you can probably draw it.
• Maybe finding a Northwestern Blot image may be a bit difficult, but there should be tons of Western Blotting photos and Northern Blotting photos that are free. The Western Blotting wiki page has some pictures maybe for you to use.

Technique specifics

• It’s a little ambiguous how the RNA binds to the proteins exactly. Could you elaborate upon that?
• It’s a bit hard to follow the whole protocol as it’s all lumped together. Also, there are a couple paragraphs here that are only 2 sentences but each paragraph should at least be 3-4 sentences. Possibly split it up more evenly at the appropriate locations to look tidier.
• There are a lot of sentences with no citations. It may be beneficial to use other citations too to make sure the article cited at the end of the whole section isn’t the only protocol that follows these steps.
• I’m not sure if the technique summary is necessary. Instead, providing a picture protocol would be better for the overall viewer. You can look at the Western Blotting article picture for ideas or possibly even use that and add onto it.

Applications

• “The blot COULD be analyzed…” Might be better if you said “can”
• The sentences are a bit choppy here and don’t connect well. Rewording it to have a better flow would be nice.
• It may be better to have actual applications included. The autoradiography, analyze, and interpret part is part of the technique and chromatography isn’t an application, but what you do after the blot is finished. It’d be nice to see what exactly researchers do with these blots and how they have applied it to their studies.
• Paragraphs shouldn’t end without a citation.

Advantages and disadvantages

• Applications and advantages seem almost the same. Any other advantages?
• Some commas can be added to the disadvantage section.
• Paragraphs shouldn’t end without a citation.

Overall, looks like the article is going in the right direction! Adding a bit more information overall would help the article greatly. Hope my review is helpful! Martinhyou (talk) 16:38, 14 April 2014 (UTC)

Thank you for the feedback martinhyou, your comments are helpful. We'll start addressing them in the next few days. Much appreciated.Msmrugby (talk) 03:30, 16 April 2014 (UTC)

Regarding your comment about the image in the origin section, I have uploaded an image of a Western Blot and also removed the previous link statement.Msmrugby (talk) 02:09, 22 April 2014 (UTC)

The Technique Specifics and Applications sections have been revised to improve the flow per your suggestions. Thanks again.Msmrugby (talk) 02:27, 23 April 2014 (UTC)

REVIEW BY KATHERINE BARNHILL

• In the lead section, I would like to see more of an overview of the entire article, it seems that the current information is more of an explanation of the technique rather than a summary of the contents of the article. Otherwise, it is clear, concise, and easily understandable. Maybe include some more wiki links to avoid potential confusion.
• Origin section-I think it would be helpful and aesthetically pleasing to have an actual image here, to give people a visual as to what you're referring to.
• I like the technique specifics section, as it gave me a nice refresher on what a northwestern blot is! However, I suggest adding more wiki links in the text to assist for understanding purposes (especially the summary items--why is there a summary??)
• Applications and advantages/disadvantages seem to have a lot of similarities, maybe consider combining them?
• The tone is clear, straight-forward and neutral; the jargon is not too technical and is easy to follow.
• The references are cited correctly and appear to be accurately applicable.
• Overall, the article is really coming together and I think it will be a great addition to the wonderful world of wikipedia information! My main suggestion is to get some images on the page and to include wiki links so that the lay person can have a more thorough understanding of the subject.

Klbarnhill (talk) 19:59, 15 April 2014 (UTC)

Thank you for the feedback Katherine. We'll be revising the article in the next few days and will certainly look to address these items. Much appreciated! Msmrugby (talk) 03:32, 16 April 2014 (UTC)

Along with Msmrugby, I would also like to thank you for your comments on the article. We are continuing to search for images to help enhance the article as a whole, however this has been one of the harder things for us to find. Tmckenne (talk) 02:17, 22 April 2014 (UTC)

We did add an image of a Western blot to the origin section and indicated that a Northwestern blot is similar looking. Also, regarding the overview, we will continue to look for ways to enhance it. Thanks again for the comments. For the technique specifics section, we are considering adding a process flow diagram but first need to create it.Msmrugby (talk) 02:21, 22 April 2014 (UTC)

Review from Keilana

Great job so far guys! Here are some suggestions for you as you go forward. Please let me know if you have any questions!

• Typically, you don't cite anything in the lead of the article, as long as it is discussed and cited elsewhere, which it typically should be.
• Nice job paraphrasing - I didn't find any plagiarism or close paraphrasing. Yay!
• The sentence referencing the SDS-PAGE image isn't quite appropriate there. It should either be an external link or you should find a free alternative image that you can put in the article.
• You should do a copyedit to check for spelling and grammar errors - Word usually does pretty okay with this.
• The second paragraph of "Technique specifics" needs a citation.
• I don't think the "technique summary" section is necessary; Wikipedia isn't really a how-to manual. If you'd like, you can put a tutorial on how to run a Northwestern on [en.wikibooks.org WikiBooks]
• The second paragraph of "Applications" needs citations, as does part of "Advantages and disadvantages"

All the best, Keilana|^{Parlez ici} 01:05, 17 April 2014 (UTC)

Thank you for the review Keilana. These comments are helpful and well look to address them. I appreciate the guidance on the image, as I've been struggling with how best to deal with this.Msmrugby (talk) 14:23, 19 April 2014 (UTC)

Hi Keilana. Just wanted to follow back up. All citations have been removed from the lead section. In addition, a copyedit was completed, and revisions were made. Regarding the Technique summary, we're looking at possibly including a process flow graphic instead of the text that is there currently. Still need to address Applications section. Thanks again for the feedback!Msmrugby (talk) 03:21, 22 April 2014 (UTC)

Just following back up on one item. The technique summary has been removed. The plan is to ultimately replace this with a process flow graphic, but this still needs to be created. Thanks again for the feedback.Msmrugby (talk) 02:35, 23 April 2014 (UTC)

Keilana, thanks for your comments on our article. All thoughts help us get it in better shape. I'm wondering why you think the technique summary section should be removed. When you look at other Wiki articles, Western blots, gel electrophoresis, etc. they go in to even more detail about how to do it then we did. I think that by explaining how to do a Northwestern blot it helps the reader understand the entire process more. Just my thoughts. Tmckenne (talk) 02:50, 23 April 2014 (UTC)

I would have to agree, the summary is useful. It's certainly not a comprehensive protocol (and shouldn't be) but gives the reader an idea of the steps taken to complete a northwestern. Additionally, including the summary allows readers to compare/contrast this technique with others. Rmiller587 (talk) 17:47, 28 April 2014 (UTC)

I agree the technique section is good. Keep in mind that the comments by any OA, even @Keilana:, are just suggestions. Some articles do go overboard with "how to" instructions, and they can become non-encyclopedic as a result. But I'd say what you have doesn't go to that level. Klortho (talk) 05:18, 3 May 2014 (UTC)

@Klortho: Hi guys, don't worry too much about this section! That was definitely just a suggestion and was more directed at keeping you from including the whole protocol. :) I really like the idea of having a graphic to show the whole process, too. Keep up the great work! Keilana|^{Parlez ici} 15:41, 3 May 2014 (UTC)

Good discussion all! We ended up including a process flow that outlines the technique steps and hopefully addresses the points made above. Thanks for the input!Msmrugby (talk) 04:07, 6 May 2014 (UTC)

Review by Crandel5425

Hey guys, I'm going to review your hard work on this article.

• In the lead is it typical to state other methods? The connection between the western and northern blots isn't out right stated. I get lost as to why they are there and felt like I was reading 3 different leads. The flow as well didn't mesh. Something like "there are two types of assays that the northwestern blot is based off, these are XX" and then explain why they are similar perhaps?
• The pictures are great, maybe a schematic of the steps required for the assay (nothing to detailed, just simple).
• Organization of the pictures - looks a bit cramped at the top with the three images there. add in a few lines to help separate the pictures from the other topics.
• In the origin section include more origin about the main topic. It seems like most of the information is driven by the southern blot. That's not to say to remove it, the focus should be back on the northwestern blot's who, what, why, when.
• A section of step-by-step (not detailed) direction of how a northwester blot is prepared - I saw someone say the opposite, but to supplement the short step-by-step heading (or pictograph) can be an external link to a public document of how a company or school executed their northwestern blot (which you already do the external link protocol section). This tells the story of how one goes about doing a northwestern blot - as we would need to know how, what, why a northwestern blot works.
• I think your external links aren't working. Also, without looking at the code, how do we input the "see also" and "external links" is that automatic like the references or manual?
• I think you found a lot of good links that were not pay-to-read.

Overall very good job, Crandel5425 (talk) 03:38, 23 April 2014 (UTC)

Thank you for the review Crandel15425. We'll work on the lead section. We received multiple comments on this, so we'll revise the structure slightly to improve clarity. Regarding the schematic, we agree. We are working on a graphical representation of the process that we plan on including. We'll be revising the organization of the pictures slightly to improve the look. The origin suggestions are noted, and this has been mentioned by others as well. It's been very difficult to find information on who created/named the Northwestern blot. The only link that we've uncovered thus far is to Southern. We'll keep looking though. We'll also take a look at the external links and see if we can fix them. Thanks for the comments!Msmrugby (talk) 12:17, 28 April 2014 (UTC)

Hi Crandel5425. Just wanted to circle back and provide an update. We've revised the images and added a process flow. I think it looks much better visually and also provides information on the technique steps. In addition, we've renamed the origin section "history" to better fit the content. We're still looking for additional relevant content to include in this section. Thanks again for the feedback.Msmrugby (talk) 04:04, 6 May 2014 (UTC)

Comments from Klortho

This is a really nice article. It is well written, has some nice figures, and has a good structure and good references. The only thing I think could use improvement is that it seems to be missing some important content. Below are some specific comments.

• In the lead, I would move "The Northwestern blot is a hybrid technique" to before you introduce the Western and Northern blots, otherwise someone reading might be confused (as I was) about why you are introducing those other techniques.
• You have some really nice images, but if you could find a cartoon representation of the process, I think that would add a lot of value to this article. It is a difficult topic to conceptualize. Something along the lines of Northern_blot#mediaviewer/File:Northern_Blot_Scheme.PNG would be great.
• I know it is late, but I am also wondering if you mightn't be able to add a little bit of content regarding the context of this technique. You might rename the "Origin" section to "History", and, in addition to how it began, describe how the technique evolved, it's popularity, other competing techniques, and whether or not it is still popular.
• Similarly, in the "Advantages and disadvantages", I am left wondering about what (if any) alternative methods there might be. Usu. advantages and disadvantages are described with reference to alternatives.

Klortho (talk) 03:49, 27 April 2014 (UTC)

Thank you for the review Klortho. We'll revise the lead section per your suggestion. Regarding the images, we agree and are trying to develop a graphical representation of the process. We hope to have it in soon. Good suggestion regarding additional content for the origin section. I think we can find some information on competing techniques and it's current relevance. Thanks again.Msmrugby (talk) 12:11, 28 April 2014 (UTC)

Klortho, thank you for your review. We are looking in to your comments. Specifically I'm going to do some research and see what, if any of the alternatives to northwestern blotting are. If I find some I will work them in to the advantages and disadvantages section. Tmckenne (talk) 21:56, 4 May 2014 (UTC)

Hi Klortho. We've revised the lead section accordingly. We've also reworked the images and added a process flow. Lastly, we reworded the origin section, as there have been multiple comments about the origin topic, and it has been extremely difficult to dig up true origin information. We're still looking for additional content on competing techniques, popularity, etc.. Thanks again for the feedback.Msmrugby (talk) 04:01, 6 May 2014 (UTC)

Review by BigA726

Hey, good job on the article so far. I do have some suggestions though.

• Your lead section follows many of the style guidelines. However, the term "Northwestern Assay" is another name for the main topic and therefore should be bolded. The sentence beginning with "A colored precipitate..." is more relevant to a procedure section later on in the article.
• Your sources seem to be in pretty good shape.
• The writing is clear and doesn't contain a lot of jargon all at once which makes it a good read.
• The Origin section should really concentrate on the origin of the Northwestern blot. I think its a good idea to start with the origin of the southern if thats where the technique originates, but almost no information is given about the origin of the topic of the article. Who first used or made the technique? When was it first made or used? How has the technique changed over time? It just feels like theres a lot of missing information in this section. Also, "When the approximate size of the protein is known, the original sample can be run on a chromatography machine to separate it by size." the "it" should be changed to something like "the proteins." for clarity. Otherwise, i feel the sections appropriately relate the information needed.
• The pictures could be spaced better throughout the article. Its very cramped looking at the top. I think the results could be at the upper right first, the picture of Edwin Southern where it is, and the apparatus used for gel electrophoresis could be in the technique specifics section.

The article seems to be written from a neutral perspective and doesn't seem biased towards or against the subject. It follows the Wikipedia guidelines well. Great Job!BigA726 (talk) 00:24, 28 April 2014 (UTC)

Thank you for the review BigA726. Your feedback is appreciated. This is a challenging topic to get origin information on or information on who specifically named it/created it. We've looked for it for weeks, and have not had any luck yet. The way it is currently explained (tracing it to Southern) is the best information that we've been able to find to date. We'll bold Northwestern Assay per your suggestion. Also good suggestion regarding images, we'll revise the placement accordingly so it presents better. Thanks again.Msmrugby (talk) 12:07, 28 April 2014 (UTC)

Not a problem! The pictures in the article look great. It looks much more spaced out. I can understand the difficulty in finding information. Otherwise, the article looks great!BigA726 (talk) 13:23, 28 April 2014 (UTC)

Hi BigA726. We've revised the images and added a process flow. I think it made a big difference, thanks for the feedback. We've also addressed the lead section comment. Still looking for additional origin information though.Msmrugby (talk) 03:56, 6 May 2014 (UTC)

Great job with the new figures. I was very impressed to see you had changed them, especially the addition of the process flow one. Your content is great too. You did a great job adding enough info about protocol to help the reader understand the overall process without making the article look like a page out of a lab manual. Again, GREAT job! Rmiller587 (talk) 01:29, 7 May 2014 (UTC)

Thanks for the kudos Rmiller587! We couldn't have done it without reviews from people like you! Thanks again! Tmckenne (talk) 01:42, 7 May 2014 (UTC)