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TRiP
>> FAQ
Below is a list of Frequently Asked Questions received by
the TRiP.
What genetic background does the TRiP use to generate their
RNAi stocks?
The TRiP stocks are in a y v; attP2 genetic background
(genotype: y[1] v[1]; P{y[+t7.7]=CaryP}attP2).
All TRiP stocks have the genotype:
y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JFXXXXX}attP2
or
y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMXXXXX}attP2,
where JF are TRiP stocks made during our pilot
project at Janelia Farm and HM are
TRiP stocks made at Harvard Medical
School.
Where are the TRiP stocks integrated into the genome?
The attP docking site of all TRiP stocks is attP2
located on the third chromosome at 68A4.
What is the eye color of the TRiP stocks?
All TRiP stocks have a wildtype appearance as the
wildtype vermillion gene is used as the selection marker
for the hairpin transformants and the attP insertions
carry the wildtype yellow gene.
Do the TRiP stocks contain "mini-white" marker?
No TRiP stocks contain "mini-white".
All TRiP stocks have the genotype:
y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JFXXXXX}attP2
or
y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMXXXXX}attP2,
where JF are TRiP stocks made during our pilot
program at Janelia Farm and HM are
TRiP stocks made at Harvard Medical
School.
What GAL4 drivers does the TRiP use to test their RNAi
lines?
Many of the TRiP lines have been tested for their
"knock-down" abilities. Since this initial testing,
however, the TRiP has entered the "production phase" where
not all lines are being tested.
The GAL4 drivers commonly used by the TRiP are listed
in the TRiP Toolbox. Some of
these drivers also contain UAS-dicer2, which enhances the
knock-down. These lines can be obtained from the BDSC.
Is it important to have UAS-dicer2 in our GAL4 line?
If the TRiP stock you are using was constructed in the
VALIUM10 vector, it is likely that UAS-dicer2 will not be
necessary to achieve maximum knock-down of your target
gene.
However, if the TRiP stock you are using was
constructed in the VALIUM1 vector, it is essential that
UAS-dicer2 be carried in the GAL4 driver you are
using.
To determine which VALIUM vector your TRiP stock
utilized please refer to the list
of available TRiP stocks.
What do I do if a TRiP stock does not give me an expected phenotype?
We welcome feedback!
If a TRiP stock does not produce an expected phenotype please email the TRiP and let us know. The construct will be removed from the collection and tested by the TRiP team. If sufficient knockdown is not observed, a new set of oligos will be designed and a new construct generated to replace the TRiP stock.
Where does the TRiP have their hairpin containing constructs injected?
Injection of TRiP hairpin containing constructs is performed
by Genetic Services, Inc. (GSI).
How do I acknowledge the TRiP?
If use of any of the TRiP stocks or reagents results in data that is included in a manuscript for publication, the TRiP requests that a version of the following statement be included in the Acknowledgements section: "We thank the TRiP at Harvard Medical School (NIH/NIGMS R01-GM084947) for providing transgenic RNAi fly stocks and/or plasmid vectors used in this study."
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