Page Comparison

Sanger Sequencing

A.  Individual Samples (<24 samples)

• • Use a 1.5 mL microfuge tube
  • Provide at least 10 uL of sample (12-15 uL preferred)
  • Centrifuge your sample before dropping it off at DSF
  • Write your order number and sample NUMBER legibly on the cap like this:

• • cap             

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B.  96 Well Plate (>24 samples)

• • Use a 96-well PCR (i.e. conical well) plate (DSF uses Eppendorf twin-tec 951020600)
  • Provide at least 10 uL of sample (12-15 uL preferred)
  • Seal your plate with adhesive foil (DSF uses VWR 60941-074)
  • Centrifuge your plate before dropping it off (you can use our Sorvall Legend X1)
  • Write your order number legibly on the plate and foil
  • Put your samples in order vertically, starting with well A1, as shown below:

                    Image Removed

PCR Cleanup

 A.  Individual Samples (<24 samples)

• • Use a 1.5 mL microfuge tube
  • Provide at least 20 uL of unpurified PCR reaction
  • Centrifuge your sample before dropping it off at DSF
  • Write your order number and sample NUMBER legibly on the cap like this:

Image Removed

• • Provide at least 3 uL of primer per sample in a 1.5 mL microfuge tube
    
  • Primer should be provided at 10 uM concentration  
  • Write the order number AND primer name legibly on the cap like this:

• • .                                      

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B.  96 Well Plate (>24 samples)

• • Use a 96-well PCR (i.e. conical well) plate (DSF uses Eppendorf twin-tec 951020600)
  • Provide at least 20 uL of unpurified PCR reaction
  • Seal your plate with adhesive foil (DSF uses VWR 60941-074)
  • Centrifuge your plate before dropping it off (you can use our Sorvall Legend X1)
  • Write your order number legibly on the plate and foil
  • Put your samples in order vertically, starting with well A1, as shown below:

                    Image Removed

• • Provide at least 3 uL of primer per sample in a 1.5 mL microfuge tube
    
  • Primer should be provided at 10 uM concentration  
  • Write the order number AND primer name legibly on the cap like this:

                                                 Image Removed

What NOT to do!

PLEASE NOTE:  

• Do NOT pool like samples in one tube! # of sample tubes must match # of requested sequencing reactions (1:1)
  
• Do NOT write your sample NAME on the lid of the microfuge tube
• Do NOT submit primer separately for Sanger sequencing orders (okay for PCR Cleanup only)
  
• Do NOT use 0.5 mL microfuge tubes for samples or primers
• Do NOT submit non-centrifuged samples.  If there is at least 4 uL at bottom, we will use as is.
• Do NOT submit orders of >48 samples in individual tubes  
• Do NOT use standard aluminum foil to cover your 96 well plates

Thank you for your cooperation!