As part of a collaboration with NASA on the Biomolecular Sequencer project, Earth-bound researchers a bicoastal team at UCSF and Weill Cornell Medical College analyzed the sequencing data from space and compared it to identical samples sequenced
on the ground.
In 2001, the group sequenced
330 small RNAs using traditional Sanger sequencing to identify 55 microRNAs.
Rubin's group sequenced
DNA from the same Croatian fossil using a method that makes many copies of DNA fragments by putting them into bacteria.
To contrast these genes, none of the 16 variable sites in MLP had an MAF > 5% in the sequenced
population (n = 90 samples), and therefore, none can be considered common in the population.
An independent laboratory has tested these conditions on an additional 10 targets in the size range 250-530 by and successfully sequenced
PCR products purified by using a MultiScreen PCR plate (Millipore Corp., Bedford, MA, USA) were sequenced
with a DNA sequencing kit (Big Dye Terminator Cycle Sequencing V2.0; PE Biosystem, Courtaboeuf, France) and subjected to electrophoresis with the 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).
The Max Planck Society's decision to fund the project is based on the analysis of approximately one million base pairs of nuclear Neandertal DNA from a 38,000-year-old Croatian fossil, sequenced
by 454 Life Sciences and reported in the aforementioned Nature paper.
The team has already sequenced
segments of 500 bases, he says.
The resulting PCR product was directly sequenced
using a SequiTherm Long-Read kit (Epicenter Technologies, Madison, WI), infrared-labeled primers S-D-Bact-0341-a-S-17 and S-*-Univ-0907-b-A-20 (LI-COR Corp., Lincoln, NE), and an automated DNA sequencer (model 400L; LI-COR Corp.).
DNA fragments from C/preM and NS5 genes amplified by RT-PCR assays employing primers WN212/WN619c (5'-TTGTGTTGGCTCTCTTGGCGTTCTT-3'/5'-CAGCCGACAGCACTGGACATTCATA-3') and WN9483/ WN9794c (5'-CACCTACGCCCTAAACACTTTCACC3"/5'-GGAACCTGCTGCCAATCATACCATC-3') were sequenced
. DNA products were analyzed on a 2% agarose gel, and the bands of the correct predicted size were excised and purified with the Gene Clean Kit (BIO 101, Carlsbad, CA, USA).
The variable body temperature is one of several traits that the NHGRI panel notes as making the lesser hedgehog tenrec an interesting contrast to the other Afrotherian about to get sequenced
, the elephant.
Current technologies are able to produce the sequence of a mammalian-sized genome of the desired data quality for $10-50 million; the goal of this initiative is to reduce costs by at least four orders of magnitude, so that a mammalian-sized genome could be sequenced
for approximately $1,000.