Preparing for Success in the Laboratory.pdf

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1、11Preparing for Success inthe LaboratoryPhillip P. FranciskovichThe Project . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2If You Dont Define the Project, the Project Will DefineYou . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

2、 . . . .2Which Research Style Best Fits Your Situation? . . . . . . . . . .2Do You Have the Essential Resources? . . . . . . . . . . . . . . . . . .2Expect the Unexpected . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3What If Things Go Better Than Expected? . . . . . . . . . . . . . .4W

3、hen Has the Project Been Completed? . . . . . . . . . . . . . . .4Was the Project a Success? . . . . . . . . . . . . . . . . . . . . . . . . . . .4A Friendly Suggestion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4The Research . . . . . . . . . . . . . . . . . . . . . . . . . . .

4、. . . . . . . . . . . . . .5Are Bad Data a Myth?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5What Constitutes a Successful Outcome? . . . . . . . . . . . . . .5What Source of Data Would Be Most Compelling? . . . . . .5Do You Have the Expertise to Obtain These Types of Data? . . .

5、. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5What Can You Do to Maximize the Reliability of YourData? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6Are You on Schedule? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7Whic

6、h Variables Require Controls? . . . . . . . . . . . . . . . . . .7The Roles of Reporting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8The Rewards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9Bibliography . . . . . . . . . . . . . . . . . . . . . .

7、. . . . . . . . . . . . . . . . . . . .9Molecular Biology Problem Solver:A Laboratory Guide. Edited by Alan S. GersteinCopyright 2001 by Wiley-Liss, Inc.ISBNs: 0-471-37972-7 (Paper); 0-471-22390-5 (Electronic)THE PROJECTIf You Dont Define the Project, the Project Will Define YouOne of the first and

8、toughest questions researchers must answer to foster success in the lab is: What do I have to accom-plish? This requires you to understand your purpose to the larger task at hand. If your research is self-directed, the answerwill most likely differ from that for someone working as part of a team eff

9、ort or answering to an immediate supervisor or experimental designer. Ask them (or yourself) what the ultimategoals are and what constitutes a successful outcome. Establishwhat constitutes compelling evidence. By projecting ahead itbecomes much easier to characterize the nature of the desiredoutcome

10、.This approach allows for problem reduction and reasonabletask planning. The greatest mistake one can make is to reacthastily to the pressures of the research by jumping in unprepared.By starting with the big picture, the stage is set for working back-ward and reducing what might otherwise appear to

11、 be a dauntingundertaking into a series of reasonably achievable tasks. This exercise also establishes the criteria for making the many deci-sions that you will face during the course of your work.Which Research Style Best Fits Your Situation?Certain decisions will have a profound impact on the natu

12、re and quality of your efforts. Some scientists favor deliberate attention to detail, careful planning and execution of each ex-periment. Others emphasize taking risks, skipping ahead andplunging in for quick results. You might want to consider whichapproach would best satisfy your superior(s) and c

13、olleagues.Eachof these “styles” has its benefits and risks, but a well-balancedapproach takes advantage of each. Sometimes it is essential toobtain a quick answer to a question before committing a sub-stantial amount of time to a more diligent data-collecting phase.Be sure everyone involved is in ag

14、reement and then plan youractivities accordingly.Do You Have the Essential Resources?Evaluate your circumstances with a critical eye. Look at yourschedule and that of your collaborators.Is everyone able to devotethe time and energies this project will demand with a minimum of distractions? Check you

15、r facilities; do you have access to thematerials and methods to do the job? Do you have the support 2Franciskovichof the decision-makers and budget managers for the duration ofthe work?Whether or not problems were uncovered, share your findings with your director and collaborators; the objective of

16、this phase is to build a consensus to proceed with no furtherchanges.Expect the UnexpectedHow flexible is your research plan? Have you allowed yourselfthe freedom to adapt your strategy in light of unanticipated out-comes? This happens frequently and is not always bad news.Unexpected results might r

17、equire slowing down the process orstopping altogether until a new path can be selected. Perhapswhole elements of the work might be skipped. In any case youshould plan on midcourse corrections in your schedule. You cant always eliminate these redirections,but if you plan for them,you can avoid many u

18、nnecessary surprises.There are likely to bemultiple paths to the desired outcome. If the unexpected occurs,consider categorizing problems as either technical or global.Tech-nical problems are usually procedural in nature.The data obtainedare either unreliable or untenable. In the former case the gat

19、her-ing of data may need to be repeated or the procedure optimizedto the new conditions in order to increase data reliability. In the later case the procedure may prove to be inadequate and analternative needs to be found. A global problem is one in whichreliable data point you in a direction far re

20、moved from the original plan.Technical problems are ultimately the responsibility of the prin-cipal investigators,so keep them informed.They might provide thesolution, or refer you to another resource. Sometimes these prob-lems can take forever to fix, so an upper limit should be agreedupon so that

21、long delays will not be an unpleasant surprise to theother participants. Delays can be the source of much resentmentamong team members but should be considered an unavoidableconsequence of research.Global problems might require more drastic rethinking.The challenge for the investigator is to decide

22、what constitutes a solvable technical glitch and what comprises a serious threat to the overall objectives. Experience is the best guide. If you have handled similar problems in the past, then you are the best judge. If you havent, locate someone who has. In any casecommunicate your concerns to all

23、involved parties as early as possible.Preparing for Success in the Laboratory34FranciskovichWhat If Things Go Better Than Expected?How can you use good fortune to your best advantage? Mostresearch triumphs are a blend of good times and bad.When goodthings happen during the course of your work,you ma

24、y find your-self ahead of schedule or gaining confidence in the direction ofyour efforts. If you find yourself ahead of schedule, think aheadand use the extra time to stay ahead.More often than not there will be subsequent phases of thework for which too little time has been allocated. Start the nex

25、tstep early or spend the time to address future problem areas of the plan. If the nature of the success you have achieved is toeliminate the necessity for some of the future work planned, youmay be tempted to skip ahead. Such a change would constitute asignificant departure from the original plan, s

26、o check with yoursuperiors before proceeding on this altered course.When Has the Project Been Completed?A project will end when the basic objectives have been met.This view of the end is comforting in that you have specific objectives and a plan to achieve them, but disconcerting if theobjectives ch

27、ange for reasons described above. If changes werecontrolled, discussed and documented throughout, endpointsshould still be easy to identify. This is another reason why it is so important to establish a written consensus for each deviationin the plan.Was the Project a Success?If you stuck to your ori

28、ginal plan and encountered no problemsalong the way, you were lucky. If problems required you to adaptyour thinking, then real success was achieved. Remember, truefailures are rare.The process of conducting research is one of con-stant evolution. If you have maintained an open mind and basedyour dec

29、isions on the facts uncovered by your work, your effortswere successful.A Friendly SuggestionIf you are a new investigator or otherwise engaged in researchthat is new to you,take a lesson from the “old-timers.”Its not thatthey have all the answers,its just that they know how to ask betterquestions.

30、They have had numerous opportunities to make theirown mistakes, and if they have been successful, it is because theyhave learned from them.Preparing for Success in the Laboratory5THE RESEARCHAre Bad Data a Myth?Data are the medium of the scientific method, and can neitherbe good or bad. Data are the

31、 answers to the questions we pose,and it is the way we pose these questions that can be good or bad. Data could have intrinsic values: indeterminate, suggestive,or compelling in nature.Poorly posed questions often lead to inde-terminate results, while exquisitely framed questions more oftenlead to c

32、ompelling data. Therefore the secret to good research isin its design.What Constitutes a Successful Outcome?The answer to this question requires another:What are the spe-cific objectives of your work? Must you produce a publication(basic research), a working model (industrial research), a reliablete

33、chnique (applications research), or a prophetic example (intel-lectual property development)?The specifications for success may vary significantly amongthese outcomes, so it might be worthwhile to verify your objec-tives with your supervisor or your collaborators.What Source of Data Would Be Most Co

34、mpelling?If the answer isnt apparent, imagine yourself presenting datain front of a group of critical reviewers.What sort of questions orobjections would you expect to hear? Answers to this question canbe gleaned from seminars on topics similar to yours and from thescientific literature.The data pub

35、lished in peer-reviewed journalshave stood up to the test of the review process and have been condensed to the most compelling evidence available to theauthor. You might also learn that the author applied an unex-pected statistical analysis to support their conclusions.Do You Have the Expertise to O

36、btain These Types of Data?Do you have access to the specific equipment, materials, andmethods necessary to perform your work? Finding access to oneof these elements can provide access to the other,as can a networkof friends and colleagues. Your desire for training might inspiresomeone to loan you th

37、e use of their equipment, along with theirexpertise.What are your options if the equipment or expertise areunavailable to you? A review of the scientific literature mightprovide you with an alternative approach. For example, if tech-nique A isnt available, the literature describing the developmentof

38、 that method will undoubtedly discuss techniques B and C andwhy they are inferior to technique A. Even if you have access totechnique A,verifying your data via technique B or C might proveuseful.What Can You Do to Maximize the Reliability of Your Data?Equipment and ReagentsIs your instrumentation wo

39、rking properly? When was it lastchecked for accuracy? An inaccurate spectrophotometer or pHmeter could affect many aspects of your research.Do you possessall necessary reagents and have you proved their potency?Have you considered your current and future sample needs?Will you employ statistical samp

40、ling in your experimental plans?You might save time, trouble, and money by analyzing your statistical sampling needs at the start of the project instead ofreturning to an earlier phase of the research to repeat a numberof experiments. How will the data be collected, stored, and ana-lyzed? How will s

41、tatistics be applied, if at all?Sample IssuesReplicatesA discussion about statistical analysis is beyond this book,but Motulsky (1995) provides practical guidance into the use ofstatistics in experimental design. Consider the use of statisticswhen determining the number of required replicates. Other

42、wise,you might find yourself returning to an earlier phase of yourproject just to repeat experiments for the purpose of statistical validation.QuantityHow much material will you require over the short and longterms? Will the source of your material be available in the future, or is it rare and diffi

43、cult to obtain? Will the physiologi-cal or chemical properties of the source change with time? What is the likelihood that the nature of your work will change,introducing new sample demands that require frequent samplepreparations?Should you prepare enough material in one episode to last theduration

44、 of your project? Sounds like a sure approach to mini-mize batch to batch variations,or is it? If the sample requirementsmake it practical to prepare an extraordinarily large amount of material, what do you know about the storage stability of the6Franciskovichprepared material? Will chemical stabili

45、zers interfere with theresearch now or in the future? Periodic control assays of materialstored over a long term might prove helpful.If the sample is subject to minimal batch-to-batch variationduring preparation, then multiple small samplings may be themost convenient approach, for this provides an

46、additional benefitof providing fresh sample.If you can verify or control for the long-term stability of yoursample, large-scale sample preparations are usually preferred,since most samples reflect the state of their source at the time thatthey are obtained.QualityGenerally speaking, samples of high

47、purity require much morestarting material, so one approach to controlling demand onsample quantities is to establish the requisite levels of purity foryour application.Many assays and experiments have some degreeof tolerance for impurities and will work well with samples thatare only moderately pure

48、. If you test the usefulness of differentsample purities in your research,you might uncover opportunitiesto reduce the required amount of sample.Are You on Schedule?You will likely be asked for precise estimates of when you planto complete your work,or for time points of certain research mile-stones

49、.The answers to the previous questions should provide youwith the big picture of the research and how the individual partscould affect one another.An accurate sense of the overall timingof the research ahead should follow.This is also a good point to search your memory, or that of a colleague who ha

50、s done similar work, to identify potential pitfalls.The goal is to eliminate surprises that tend to get you offschedule.Which Variables Require Controls?Consider the converse question: Which variables dont requirecontrols? You might have to switch sample origins, reagents,reagent manufacturers, or i