Investors in the round include OrbiMed, Pontifax Fund, RA Capital Management, EcoR1
Capital, Omega Funds, BVF Partners LP, Boxer Capital LLC, Adage Capital Management LP, and Alexandria Venture Investments.
All of the existing investors in Scholar Rock also participated in this financing round, including Polaris Partners, Timothy Springer, ARCH Venture Partners, EcoR1
Capital, The Kraft Group, Fidelity Management and Research Company, and Cormorant Asset Management.
Twenty micrograms of genomic DNA was digested with EcoR1
and separated electrophoretically on a 1.
The positive clones were confirmed via restriction analyses using EcoR1
and Pst1 enzymes.
Approximately 700 ng of DNA from each accession was double-digested by the enzymes EcoR1
and Mse1, using 5 U of each, for 18 hours at 37[degrees]C.
As in the Anolis study, we digested the genome with SphI and EcoR1
, and selected fragments from 276-324 base pairs long to recover homologous loci scattered randomly across the genome.
Capital was the lead investor in this financing, which included significant participation from Fidelity Management and Research company, ARCH Venture Partners, Boxer Capital of Tavistock Life Sciences, Partner Fund Management, Nextech Invest, as well as a number of other healthcare investors.
Oligonucleotides Purpose Sequence (5'-3') EcoR1
-forward adapter Adapter ligation CTCGTAGACTGCGTACC EcoR1
-reverse adapter Adapter ligation AATTGGTACGCAGTCTAC Mse 1-forward adapter Adapter ligation GACGATGAGTCCTGAG Mse 1-reverse adapter Adapter ligation TACTCAGGACTCAT EcoR1
primer Pre-amplification GACTGCGTACCAATTC Mse1 primer Pre-amplification GATGAGTCCTGAGTAA E-ACA Selective amplification GACTGCGTACCAATTC + ACA E-ACT Selective amplification GACTGCGTACCAATTC + ACT M-CTA Selective amplification GATGAGTCCTGAGTAA + CTA M-CTG Selective amplification GATGAGTCCTGAGTAA + CTG TABLE 3.
subtilis 168 with a length of 80 bps and its complementary sequence were synthesized and ligated into pUB110 at EcoR1
and Xba/restriction sites.
For/ Rev Klentaq1 and For/ Rev Taq were employed which during PCR was cut with two restriction enzyme EcoR1
and Sal1, cloned in plasmid PET21a and transferred to E.
by PCR amplification of spacer sequences separating a subset of copia-like EcoR1
The toxic region was restricted with EcoR1
and HindIII, and ligated in the expression vector pT7-7.