Frequently Asked Questions

• Repeat hybridization using one of the following: 
  - Decrease the melt temperature by 2°C 
  - Decrease the melt time until signal intensity becomes acceptable

• Repeat the assay on a new specimen using one of the following: 
  - Increase the hybridization time 
  - Increase the melt temperature as needed until the signal becomes adequate 
  - Wash the slide with 0.4X SSC/0.3% NP-40 at 70-73°C

• Repeat the assay on a new specimen using one of the following: 
  - Increase the temperature of 0.4X SSC/0.3% NP-40 by 2°C; continue to increase the temperature as needed 
  - Decrease the melt temperature by 2°C

-Remove cover slip and Immerse slides in 2X SSC/0.1% NP-40 at ambient temperature for 5 minutes; agitate occasionally. -Dehydrate slide through series of ethanol rinses. Air dry and reapply new counterstain 

Was the wrong concentration of counterstain used? 
-If too bright, dilute the counterstain in antifade solution before applying 

Was the counterstain too old or exposed to light for an extended period? 
-Store counterstain at -20°C, protected from light 
-Do not use counterstain past its expiration date

• Were the probes diluted inappropriately? 
  -Ensure the probe mixture was made according to protocol specifications 
  
  Were the conditions inappropriate for hybridization? 
  -Ensure incubator is at the proper temperature of 37°C 
  -Check that the hybridization buffer was added to the probe mixture and in the adequate amount 
  
  Was the wash temperature too low? 
  -To maintain the wash temperature, place no more than four slides in the wash at a time and ensure temperature of the wash solution is correct prior to washing another set 
  
  Was the wash solution stringency too low? 
  -Ensure wash solutions are made according to protocol 
  -The lower the concentration of salt, the higher the concentration of formamide and NP-40, and subsequently, the more stringent the wash

• Was slide cleaned properly prior to sample preparation? 
  -Immerse slide in ethanol, dry using lint-free paper 
  
  Were the Specimen slides used too soon prior to denaturation? 
  -Ensure slides are aged at least 24 hours at ambient temperature 
  
  Could there have been too much cellular debris on the sample? 
  -Wash cell pellet with fresh fixative three times prior to dropping slides 
  
  Were the metaphase spreads aged by baking, or do they contain cytoplasm? 
  -Increase time the slide is immersed in the denaturation solution to 10 minutes 
  
  Was the slide inadequately washed following hybridization? 
  -Ensure wash solutions were made according to procedures outlined by the package insert 
  -Make sure the wash solution is at the proper PH and temperature, remove cover slip and repeat the wash procedure 
  -Increase immerse time to 4 minutes in the 73°C 0.4XSSC/0.3% NP-40 wash 
  
  Were the wash solutions used too long or improperly stored? 
  -Ensure wash solutions containing formamide are properly stored at 4°C 
  -Discard these solutions after 7 days or after frequent use; discard all other solutions after 1 day 
  -Ensure the pH of formamide solutions are pH 7.0-8.0 
  
  Was the hybridization viewed using long bandpass filters? 
  - Reduce background light by switching to filters with smaller bandwidths or to multi-bandpass filters

• Did the slides dry too quickly during preparation? 
  -Increase the humidity used when dropping slides, or increase the temperature of the water bath. 
  -Decrease slide warmer temperature during preparation of samples 
  -Let slides dry overnight; age for at least 24 hours at room temperature 
  -Do not bake slides at high temperature 
  
  Was the specimen over-denatured? 
  -Adhere to directions on package insert to make denaturation solution 
  -Prior to immersing the slide, ensure temperature of the solution is at 73°C, decrease temperature to 72°C 
  -Decrease the time of slide denaturation process by 1-3 minutes

We can accept RNA, DNA, peripheral blood, bone marrow, and FFPE samples.

Our requisition form can be found here along with other protocols and information.

Our specimen requirements can be found here along with other protocols and information.

Yes. If you are shipping RNA/DNA to US from outside of the US you are required to include some Customs documentation with your shipment. To prepare this document, we require the following information from you: your name, mailing address with postal code, e-mail address, phone number, name of courier you are using (ie. FedEx), the number of packages being sent and the approximate weight/size; and a description of what you are sending (non-hazardous, non-toxic, non-infectious RNA/DNA only; please specify the organism the RNA/DNA is from and the number of tubes you are sending with approximate volume of each). Please indicate the samples as having zero value. If you are shipping within the US, no such documentation is required.

Turnaround time is variable based on the type of testing ordered and the number of samples requested. A cytogenetic or FISH test typically takes around 7 days per sample. For more detail on molecular tests with turnaround time details click here.

Questions can be emailed to info@empiregenomics.com or we can be called directly at (716) 856-3873.