We are looking into upgrading to the 8-capillary AB 3500 Genetic Analyzer. We currently have a 16-cap 3130xl, but it's getting discontinued. We are a small core facility that sequences various sample numbers each week; from 2 samples one week, to 192+ another week.
1) Does anyone have experience using the 8-capillary sequencer that would be willing to share any cost-saving tips? I'm concerned about the RFID pouches that the 3500 requires; we normally stretch our reagents and purchase off-brand products to keep our costs low.
2) Can all the "expiration warnings" be overrode or ignored?
3) Can someone tell me which "expiration" dates the instrument can be overrode and if there are any things that the instrument will not let you move forward using? Ex. Can I use a polymer pouch that has been on the instrument for 4 weeks, or will it lock me out of using it?
4) Will a full plate of sequence samples degrade by the time the run is complete? Almost 25 hrs to complete a full 96-well plate. I'm concerned about C-Tails.
I'd really appreciate any insight from someone who physically uses the 3500 (as opposed to Thermo customer support who tells you the best-case scenarios). TIA!
Dear Lisa McDougall
Technologies for next-generation sequencing (NGS) are comparable. While the Sanger technique only sequences a single DNA fragment at a time, NGS is massively parallel and may sequence millions of fragments at the same time per run. This high-throughput method allows for the simultaneous sequencing of hundreds to thousands of genes.
Note:
The draft sequencing indicated that the human genome had much fewer genes than predicted. Previously, counts up to and exceeding 150,000 genes were predicted, but the draft sequencing revealed that there were about 30,000-40,000 human genes at most.
A DNA fragment-containing sample is inserted into the capillary. This is accomplished by immersing the capillary and an electrode in a sample solution and temporarily applying an electric current. After injecting the sample, the electric field is reapplied to force the DNA fragments through the capillary.
In molecular biology, sequencing is used to investigate genomes and the proteins they encode. Researchers can use sequencing data to find changes in genes, connections with illnesses and phenotypes, and possible therapeutic targets.
The HGP employed two key approaches: Expressed Sequence Tags (ESTs) — This method identifies all genes that are expressed as RNA. Sequence Annotation — This method included blindly sequencing the whole genome (coding and non-coding) and afterwards assigning functions to the various sections.
Kind Regards
Qamar Ul Islam
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