spinvorti.blogg.se - Macvector trim chromatogram

#MACVECTOR TRIM CHROMATOGRAM SOFTWARE#
#MACVECTOR TRIM CHROMATOGRAM FREE#

Low to moderate specific binding at the 3′ end (avoid high GC content to prevent mispriming).Absence of significant hairpin formation (>3 bp).

A melting temperature (Tm) in the range of 50 C to 65 C.

It is important that a primer has the following characteristics: One of the single most important factors in successful automated DNA sequencing is proper primer design. The SP6long primer is four bases longer (so check for compatibility with your vectors) but works well for large templates when the shorter SP6 primer fails.

polyA(CGT) 5′ AAAAAAAAAAAAAAAAAAAAAAAAAC/G/T 3′įor large templates such as BAC’s, PAC’s and cosmids which can have higher levels of impurities, we recommend the use of the SP6long primer instead of the standard SP6 primer, which has a marginally low Tm.

polyA-T 5′ AAAAAAAAAAAAAAAAAAAAAAAAAT 3′.

polyA-C 5′ AAAAAAAAAAAAAAAAAAAAAAAAAC 3′.

polyA-G 5′ AAAAAAAAAAAAAAAAAAAAAAAAAG 3′.

polyT(ACG) 5′ TTTTTTTTTTTTTTTTTTTTTTTTTA/C/G 3′.

polyT-A 5′ TTTTTTTTTTTTTTTTTTTTTTTTTA 3′.

polyT-C 5′ TTTTTTTTTTTTTTTTTTTTTTTTTC 3′.

polyT-G 5′ TTTTTTTTTTTTTTTTTTTTTTTTTG 3′.

We supply the following standard primers (most at 4 uM): Samples MUST be organized in the 8-well dimension in plates, with samples running from 01A, 01B, 01C, etc. Organize reactions to avoid gaps where possible. Please give samples and primers reasonably short names, avoiding more than a combined length of 23 characters. For shipped samples it is especially important that the sealing method be secure against leakage, and samples packed to avoid crushing. Sealing with most tapes (clear & metallic) WILL result in leakage unless the samples are dried down. Samples should be securely sealed with strip caps. Volumes for templates and primers should reflect that we use 1.4 and 1.0 ul respectively for each reaction. Reactions should be organized in 96-well PCR-type plates, or in 8-tube strip tubes such as these:ĭO NOT use flat-bottomed plates as they are difficult to pipette from.

#MACVECTOR TRIM CHROMATOGRAM FREE#

High throughput reactions receive a lower price because reactions are prepared using multichannel pipettors, the resulting chromatograms are not edited, and free redo reactions are limited to facility errors, not user errors. Direction for using the portal may be found here: End User Instructions.Įxternal Users:Samples may be dropped off as above, or shipped to DNA Sequencing Facility, University of Chicago, KCBD rm 1230H, 900 E 57th St, Chicago, IL, 60637īilling questions may be directed to High-Throughput Sequencing: We require a minimum of 12, but preferably 24, reactions to be provided for this service. 57th) hallway freezer outside room 1230H (north end of east corridor under the hanging shark), or at the Cummings Life Science Center 6th floor hallway (room 604, just right of the elevators) as soon as possible after submitting your request on the DNA Order Portal. Internal Users: Please drop off samples at the KCBD (900 E. Our turnaround time is 1 business day from receipt of samples (multi-plate orders may be slightly longer).įor high-throughput sequencing (24 reactions minimum) or genotyping, custom services, such as complete sequencing projects, primer design and synthesis, etc, please call us at 77. We are committed to giving every user satisfactory sequence. Users are guaranteed satisfactory sequence from their orders even if it requires resequencing new template DNA (there is a 24 reaction limit), or the failed reactions are free.Ĭlick here for our redo policy.

#MACVECTOR TRIM CHROMATOGRAM SOFTWARE#

For chromatogram editing, software is available for Macintosh and Windows platforms (see below). We also run user’s reactions, and genotyping (fragment analysis) samples. Users provide DNA (plasmid, phage or PCR product) at a standardized concentration and custom primers if necessary, and the Facility performs cycle sequencing reactions using fluorescent dye terminators, runs the capillaries, acquires the data, and provides the sequence as chromatogram and text files.