I stink at research!

[WindChaser]

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Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

 

[supermintyfresh]

Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

Yeah Westerblots and Immunostaining can get tricky. The key is to take your time and to follow the procedures correctly. Try to see other people do it in action, rather than just reading from a document, as sometimes there are techniques that aren't written down.

Also there are Youtube videos, surprisingly, on many lab techniques so check those out! Best of luck. Don't let research intimidate you, just have fun with it!

 

[Nimalin]

I hear you-- I tried out bench research for several semesters in college and was horrible at it. Part of the reason was I simply didn't like it, and the longer I stuck with it the more miserable and unsuccessful I became. I decided basic science research simply wasn't for me and joined a psychiatry lab instead. It was a lot more fun. I got to work with people and play with databases. I was happier with my work and more productive. I'm not saying you should give up if you're struggling, but just keep an open mind there are many ways to become involved in research if you decide benchwork isn't your cup of tea.

 

[Omppu27]

Yeah Westerblots and Immunostaining can get tricky. The key is to take your time and to follow the procedures correctly. Try to see other people do it in action, rather than just reading from a document, as sometimes there are techniques that aren't written down.

Also there are Youtube videos, surprisingly, on many lab techniques so check those out! Best of luck. Don't let research intimidate you, just have fun with it!

Yes. Watch someone else run the gel. The first time I started running gels I didn't get any bands for two straight weeks because I think I was doing the transfer wrong... once I watched a grad student do it again, I was perfectly fine.

 

[Penguin180]

It was difficult for me too when I started, but it takes some practice around what you do; and once you relax more things get better and better
when in doubt, it helps to ask more questions early on; and make a visual map/plan out more in advance 🙂 and most TAs/teachers understand and will help you

 

[DrEnderW]

I think a lot of us have been there with some experiment or protocol. Anything with the phrase "immuno" in it is pure witchcraft and things just don't turn out sometimes. As others have said, ask to watch someone run through the protocol again. The smallest little tips and tricks can really make a big difference. You'll get it.

 

[BlueLabel]

Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

Are you using a pre stained ladder? Does it seem to resolve well? What kind of transfer are you doing wet, dry, semidry? Are you ponceau staining the blot and coommassie staining the gel after transfer to check efficiency? Are you using ECL or fluorescent secondaries? If you're careful and doing it under experienced supervision you should be getting something. In what way are your results bad? No bands? Bubbles? Low signal?

 

[BlueLabel]

Also you need to keep in mind that all primary antibodies are not created equal. In fact some brands (Santa Cruz I'm looking at you) really stink. Check to see if others in your lab have had success with that primary. Other things to consider: check the manufacturer info for the antibody and use the maximum recommended primary concentration if you're getting a weak signal. Also, if you're in a big group and using lab stock buffers (running, transfer, TBS) try preparing them freshly yourself, being extremely careful with pH. This doesn't apply if you're using commercially available buffers, only if ya'll are mixing them in lab.

 

[Mr Avante]

Hehe. Your friend is correct. You are doing things right, but wrong. Let me explain:

You follow the protocols correctly but you lack technique. At my lab, undergrads are always efficient in following commands, but they are never aggressive with their analyses. 99% of the time it is due to the fear of breaking equipment or, naturally, ruining the experiment.

!!🙂

 

[TriagePreMed]

Ahh yes, I was a total ***** when I started. It took like 4 months for me to "get it" and through that time the post-doc would get more and more pissed at me. You just gotta keep going, but you also have to evaluate what is wrong with what you're doing, which may not be because of you. I've had situations where I have to add a primer diluted 1:20 (as was indicated to me) and never worked until I accidentally did a 1:10 dilution of it and the bands came out beautiful.

I recommend you power through this until somehow your motor skills click with what's going on.

Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

 

[Reckoner]

Also you need to keep in mind that all primary antibodies are not created equal. In fact some brands (Santa Cruz I'm looking at you) really stink. Check to see if others in your lab have had success with that primary. Other things to consider: check the manufacturer info for the antibody and use the maximum recommended primary concentration if you're getting a weak signal. Also, if you're in a big group and using lab stock buffers (running, transfer, TBS) try preparing them freshly yourself, being extremely careful with pH. This doesn't apply if you're using commercially available buffers, only if ya'll are mixing them in lab.

My old lab tried a Santa Cruz primary that labeled everything except the protein we were looking for. That was a wasted two months...

 

[WindChaser]

Thank you for all the advice and words of encouragement!

Yeah Westerblots and Immunostaining can get tricky. The key is to take your time and to follow the procedures correctly. Try to see other people do it in action, rather than just reading from a document, as sometimes there are techniques that aren't written down.

Also there are Youtube videos, surprisingly, on many lab techniques so check those out! Best of luck. Don't let research intimidate you, just have fun with it!

I'll try Youtube. Never thought of that before. Thanks!

I hear you-- I tried out bench research for several semesters in college and was horrible at it. Part of the reason was I simply didn't like it, and the longer I stuck with it the more miserable and unsuccessful I became. I decided basic science research simply wasn't for me and joined a psychiatry lab instead. It was a lot more fun. I got to work with people and play with databases. I was happier with my work and more productive. I'm not saying you should give up if you're struggling, but just keep an open mind there are many ways to become involved in research if you decide benchwork isn't your cup of tea.

I definitely LOVE bench research. It would be a dream come true if I was actually GOOD at it though. 🙁

Also you need to keep in mind that all primary antibodies are not created equal. In fact some brands (Santa Cruz I'm looking at you) really stink. Check to see if others in your lab have had success with that primary. Other things to consider: check the manufacturer info for the antibody and use the maximum recommended primary concentration if you're getting a weak signal. Also, if you're in a big group and using lab stock buffers (running, transfer, TBS) try preparing them freshly yourself, being extremely careful with pH. This doesn't apply if you're using commercially available buffers, only if ya'll are mixing them in lab.

So is it bad if our whole lab continuously uses the same buffer without changing it? The Post docs say it's fine, but is it worth using a new batch? Thanks for your advice!

What usually goes wrong is that nothing shows up and if something does show up (bands), then it's not the right length or it's way too faded.

Hehe. Your friend is correct. You are doing things right, but wrong. Let me explain:

You follow the protocols correctly but you lack technique. At my lab, undergrads are always efficient in following commands, but they are never aggressive with their analyses. 99% of the time it is due to the fear of breaking equipment or, naturally, ruining the experiment.

!!🙂

Is it necessary to be "aggressive" with my technique? Thanks!

Ahh yes, I was a total ***** when I started. It took like 4 months for me to "get it" and through that time the post-doc would get more and more pissed at me. You just gotta keep going, but you also have to evaluate what is wrong with what you're doing, which may not be because of you. I've had situations where I have to add a primer diluted 1:20 (as was indicated to me) and never worked until I accidentally did a 1:10 dilution of it and the bands came out beautiful.

I recommend you power through this until somehow your motor skills click with what's going on.

So there comes a point where everything instantly clicks right? That makes me feel so much better! Although having done this for almost a year....I hope I'm just a late bloomer...

 

[hockey40]

I would say some things to keep in mind are 1) each protocol can have its "intricacies" i.e. you'll need to do some trial and error before you find out what works and 2) develop some good trouble shooting skills. For example, I had the hardest time getting an endonuclease to cut a plasmid I was working with for two weeks before I went back and did some genomic research on the DNA strand. Turned out there was dam methylation and I needed to transfer the plasmid into a dam- strain of E. coli. Long story short, don't give up and don't be afraid to dig a little deeper. The PI will be impressed.

 

[BlueLabel]

So is it bad if our whole lab continuously uses the same buffer without changing it? The Post docs say it's fine, but is it worth using a new batch? Thanks for your advice!

What usually goes wrong is that nothing shows up and if something does show up (bands), then it's not the right length or it's way too faded.

There are many possible reasons for your problems. I'm not sure quite what you mean about the buffer but for troubleshooting purposes I'd make everything fresh. Also, ask around about the primary antibodies. If you're having a lot of trouble, what I'd advise is asking someone more experienced (not nec a post doc, ideally a friendly grad student) to either walk you through the protocol step by step or observe while you do it like you usually do, and point out your errors.

As for your difficulty with PCR, this technique is much easier to troubleshoot because there are fewer steps. Your problem probably has to do with inconsistent pipetting

 

[APrioriAporia]

Try brinigng this up to your postdoc and talk to him abt suggestions. Admit that you may need more procedural help, and see what he suggests. I would suggest that you ask if you could do the exact same protocol at the exact same time with him (two runs), so that there isn't some weird small detail you need or need more practice on getiing correct.

 

[Goro]

That's why they put the "Re" in "Re-search"

Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

 

[IonClaws]

Hey guys,

I admit that I survived my first year of college by reading SDN threads... I've recently encountered a problem that I haven't been able to find a previous thread for, so here I am!

Simply put, I STINK at research. I don't know if it's just me, but I've been doing research since fall semester of last year...and I'm STILL not good at it yet. I still mess up simple protocols like PCR's or Western Blots.

The post-doc I work with says that everything I'm doing is correct....UP UNTIL WE GET OUR RESULTS. Then when the yield is too low or bands don't show at the correct place or something...he says "Oh you probably did something wrong".

I know it's my fault. Obviously if he can do it correctly, I should be able too. But what am I doing wrong? He doesn't notice anything wrong with the way I did the protocol. What can I do? How can I improve my ability to work in a research lab?

The only reason I'm so frustrated is because he mentioned to me when I joined that if I did a good job, he will give me more responsibilities and the possibility of co authorship later on was good. But every time I mess something up, I see that disappointed look on his face. I REALLY want to get better, but if he can't identify what I'm doing wrong and I certainly can't with my little knowledge, how can I get better?

I appreciate any help I can get. 🙂

Is the post-doc looking over your shoulder while you're doing all of these techniques you "suck" at? You can still get poor results with good or even great technique when doing research. I think he might be frustrated that he isn't getting the results he wants or expects, but that's a function of what you're measuring, not how good your technique is. This is a completely normal part of research, so I doubt you "suck" at it. Just sit down and talk to the post-doc and try to figure out what you can do differently to get better results. 👍

tl;dr

That's why they put the "Re" in "Re-search"

 

[ShySpliceosome]

My old lab tried a santa cruz primary that labeled everything except the protein we were looking for. That was a wasted two months...

Lol...that's pretty sad...