Q: What should I do if I don’t collect some of the data asked for?

A: From our pilot studies, we know that some facilities, for example, do not count the total number of eggs harvested.  You should submit your data even if some categories are missing.  The statistical power will not be quite as high, but your contribution will still be valuable, and you will still receive your custom analysis.

 

Q: Some of the data we normally collect is missing for some injection days. Should we exclude those days from our file?

A: No, please include all injection days, regardless of whether you are missing data.

 

Q: Should we leave out data from injections performed by new staff who are still being trained?

A: No, please include ALL data from ALL injections. There is no way to accurately define the point at which a microinjectionist is "fully trained."

 

Q: How do I submit my data?

A: Please submit your data file(s) and facility description(s) ONLY via the survey's web interface. Do not send EXCEL files or facility descriptions to an individual person.

 

Q: We don’t normally count the pups at birth.  We usually wait a few days, so we may be missing some that die and are cannibalized.  Should we still submit the number of pups produced?

A: Yes.  Regardless of how you count the pups, you should submit the number.

 

Q: Sometimes we do not implant all of the embryos that survive the injection process.  For example, we occasionally culture a few of them to the blastocyst stage to test our culture conditions, and we do not implant these.  Should we include these in the number of eggs injected?

A: Yes, but please record this type of incident in the “Incidences” column of the Excel file.

 

Q: If a donor female is not plugged, we do not harvest eggs from her.  Instead, we put her back in the colony to be used for a future injection.  Should we include such females in the number of donors superovulated?

A: No, you should only count the females from which you actually harvest eggs.  (See description of the “Donor Females Superovulated” column.)

 

Q: Different microinjectionists in my facility have different skill levels and therefore different yields.  Is it possible to add a column indicating who did each injection?

A: We debated this question quite a bit while designing the survey.  We finally decided not to distinguish between operators for 2 reasons: (1) variation due to the person doing the work was an important part of the overall variability, and we wanted the overall results to reflect this; (2) we wanted to keep the survey as simple as possible to encourage more participation.  Several other categories of data were considered and rejected as well.  Note, however, that you can include this distinction in the “Incidences” field.

 

Q: How should I distinguish between the two types of DNA constructs?  Some constructs we’ve injected are intermediate in size between normal plasmids and normal BACs.

A: The upper limit on the size of plasmid-based transgenes is about 30 Kb.  Therefore, you should classify anything larger than that as a genomic-based construct.

 

Q: Why aren’t you collecting data on the percentage of transgenic pups that express the transgene?  This would be very valuable to know!

A: We all agreed that this could be a very informative statistic to collect.  However, most facilities do not collect this data, and if they do, it may be defined in different ways (e.g., protein versus mRNA).  In addition, most of the time, the customer collects this data, and we did not want to depend on outside data that may or may not be reported faithfully to the facility.

 

Q: Shouldn’t you ask for the number of pups weaned, since this will reflect the ability of the facility to keep pups alive and affects the number of transgenic pups produced?

A: We debated this question quite a bit also.  The problem is that some facilities genotype before weaning and some genotype after.  Those that genotype before weaning often euthanize wildtype pups before (or during) weaning, if they receive the genotyping results in time.  Also, some facilities genotype pups that die before weaning, and some don’t, so we decided there were too many differences in the way this data would be reported.  As in many other cases, this decision was driven by the desire to maximize the number of facilities that could supply data for all categories.

 

Q: We know from our own studies that the type of media used can affect the yields of DNA microinjection.  Why didn’t you include a category for this?

A: We decided not to gather this data for the same reasons we did not ask people to identify which microinjectionists performed the work.  Also, virtually all facilities use one of only a few well-studied types of media, and these are unlikely to be a large source of variability.  Therefore, we felt the value of this variable was outweighed by the desire to keep the survey as simple as possible.

 

Q: I’m worried that our facility will look bad compared to other larger facilities.  How do I know my data will be kept confidential?

A: Your original data file will be seen by only two people, Dr. Lluis Montoliu, the President of the ISTT, and Thomas Fielder, who originally proposed the idea of this survey to the ISTT.  You have our word that your data will be stripped of all identifying names before being included in the analysis.  However, we have to have some way of matching the data to the facility so we can provide the custom analysis.  We considered other ways of dealing with this, but having a human view the original files is the best way to ensure the integrity of the data.  Remember that, even if your facility is having a difficult time performing at the level you would prefer, perhaps due to changes in personnel or other factors beyond your control, your input is still valuable, and the custom analysis will allow you to see exactly how you measure up.  It will also give you an “industry standard” to aim for, so you can tell what improvements are necessary (e.g., is the problem a lack of pups or low egg yields). 

 

Q: I know that our yields are lower than the averages reported by other facilities just by reading the Tg-L.  I don’t need a custom analysis to tell me this, and my data would skew the results.  Why don’t you just collect data from the top 5 or 10 facilities in the world?

A: This is a very good question!  In fact, we are considering asking certain facilities to submit more categories of data to see if we can identify a set of factors that result in the best possible yields.  However, even the best facilities have a large amount of variation in their yields from strain to strain and from construct to construct (probably – we don’t know yet!).  This can result in dissatisfied clients who then spread rumors about the facility.  Therefore, describing this variability is just as important as determining the best possible outcomes.  In any case, until we do a very comprehensive survey that includes the best and worst data from a large number of facilities, we won’t really know how good the top 5 or 10 facilities really are, or even which ones they are!  Therefore, even poor yields are important to the success of this survey.

 

Q: We sometimes use the embryos collected in one session for microinjecting several different constructs. How should we enter/treat these data in the survey?

A: Presumably the number of embryos used for each construct will have been recorded separately, therefore we recommend that you split "ad hoc" the embryo donor females and the number of collected, microinjected and transferred embryos.

 

Q: Has this survey been tested to collect data correctly from different computers, browsers and platforms?

A: Yes, this transgenic survey has been tested and confirmed to function correctly in PCs (WindowsXP), MacOS (9.X and 10.X) and Linux (Ubuntu 8.X) with the most widely used WEB browsers (Internet Explorer, Firefox, Netscape and Opera). However, please note that Safari browser, in combination with MacOS 10.X, does not correctly process the javascript code that is used by the transgenic survey, therefore its use is not recommended. In case you would like to use Safari, please bear in mind that all forms need to be filled entirely before hitting the "return" button. The most recent WEB browser, Chrome, by Google, has produced some unexpected results (i.e. popup windows are blocked) and, thus, its use is also not recommended.

 

Q: The survey does not accept my newest Excel files (.xlsx, Office 2007 format). What can I do?

A: You have to save your Excel 2007 files (.xlsx) as "Excel 2003" files (.xls) in order for the system to accept your contributions.

 

Q: I have questions or comments about some details of the data I have submitted. How do I contact the people responsible for this survey on transgenic mouse production?

A: Simply send a message to tgsurvey@transtechsociety.org.