Smolke:Protocols/Sequencing

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Current revision (14:14, 7 July 2012) (view source)
m (Elim's recommended sample format)
 
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==Elim's recommended sample format==
==Elim's recommended sample format==
-
*At least 50 ng/ul, preferably >100 ng/ul for sample
+
*At least 50 ng/ul, preferably >100 ng/ul for plasmid sample
*3 uM for primers
*3 uM for primers
*10 ul plasmid and 3 uL primer for each sequencing reaction
*10 ul plasmid and 3 uL primer for each sequencing reaction
Line 8: Line 8:
*Use 8-PCR tube strips
*Use 8-PCR tube strips
**I prefer 0.5mL tubes, which give more room to label on the top. -Josh
**I prefer 0.5mL tubes, which give more room to label on the top. -Josh
 +
**Elim biopharm: When possible, please use 8-strip PCR tubes for less than 96 samples and primers, and 96-well plates for equal to or greater than 96 samples
*Tape sample to order form
*Tape sample to order form
**Or recycle the bags that primers come in, and just put the order form in the bag. -Josh
**Or recycle the bags that primers come in, and just put the order form in the bag. -Josh

Current revision

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Elim's recommended sample format

  • At least 50 ng/ul, preferably >100 ng/ul for plasmid sample
  • 3 uM for primers
  • 10 ul plasmid and 3 uL primer for each sequencing reaction
    • Submit 10 uL or 3 uL/rxn of primer, whichever is larger. -Josh
  • Use 8-PCR tube strips
    • I prefer 0.5mL tubes, which give more room to label on the top. -Josh
    • Elim biopharm: When possible, please use 8-strip PCR tubes for less than 96 samples and primers, and 96-well plates for equal to or greater than 96 samples
  • Tape sample to order form
    • Or recycle the bags that primers come in, and just put the order form in the bag. -Josh

Notes

  • If sequencing constructs with strong secondary structures, check GC rich on order form. I have found that "GC Protocol 2" often worked better for the various shRNA and miRNA sequences I have submitted, and you could specify this in the comments section on the order form. --YC
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