During this time frame, you may request your saved tubes to be used for new reactions (such as using a previously submitted sample to be run with a new primer) - simply submit an order form and any tubes that are not already in our possession, and refer to the date they were last used so we may find them easily and confirm whether enough volume is leftover.Tubes are saved for 2 weeks from the date they are run. M13F GTAAAACGACGGCCAG *Our default M13F is M13F(-20).Stock Primers (we supply the following primers so you don't have to! If you would like us to add another common primer to this list, please let us know by emailing T7 TAATACGACTCACTATAGGG.Custom Primers (you only need to provide your own primer if we do not have it in stock): provide at a 2µM concentration. only one primer can be used per reaction or else it will have a mixed sequence (this is an important difference between sequencing and regular PCR reactions).GC content of ~40-60% (high GC can cause mispriming).low to moderate specific binding at 3' end.no dimerization, significant hairpin formation, or secondary binding sites.annealing site more than 30 bp away from the target.primers should be an exact match with the template (a mismatch can result in a failed reaction).BACs & gDNA: 500-1000 ng/µL (for notes on gDNA prep, please see "Purification" above).Concentration: can be above or below the recommended ranges below (but please dilute if more than 50% above the upper limit).For genomic DNA (gDNA) ONLY: please isolate and amplify the specific target sequence prior to purification (*bacterial gDNA is the exception as it does not need to be isolated and amplified by PCR).Be sure to elute in autoclaved dH2O (not the kit buffer) - salt-containing buffers will inhibit reactions and cause poor reads.Purification: use a high-quality commercial purification kit (such as Qiagen).Volume: 8 µL per reaction (10 µL if sample concentration is low).
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