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 Home > Resources > Libraries
   VMRC-19: Yellowbelly rockcod (Notothenia coriiceps) BAC Library   

The Antarctic yellowbelly rockcod (Notothenia coriiceps) BAC library was constructed by Michael Schoenborn in Chris Amemiya’s laboratory at the Benaroya Research Institute at Virginia Mason (BRI). This library was designated VMRC-19. Erythrocytes were obtained from a wild-caught rockcod (sex unknown) from Dallman Bay in the Antarctic Peninsula by Chris Amemiya. Erythrocytes were embedded in Incert agarose and high molecular weight DNA was prepared in situ in the agarose. Generation of the library closely followed the cloning approach developed in Pieter de Jong’s laboratory (Osoegawa et al., 1998). Partial-digest EcoRI fragments from the appropriate size fraction were cloned into the pCCBACE1 vector (Epicentre Technologies). The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library was arrayed into 288 384-well microtiter dishes. The library has been arrayed into 384-well microtiter dishes and has subsequently been gridded onto 22 x 22cm nylon high-density colony filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones (equivalent to 48 "384-well" dishes), stamped in duplicate. Analysis of 120 randomly selected BACs via pulsed field gel electrophoresis of NotI-digested DNA indicated that the average insert size was around 138 kb, whereas the median size was ~ 155 kb. The BAC library construction was funded by NIH grant HG02526 (Chris T. Amemiya, BRI) under the auspices of the NIH Resource Network.

Segment

1

All

Vector

pCC1BAC

pCC1BAC

Restriction Enzyme

EcoRI

EcoRI

DNA Source

N/A

N/A

Plate Numbers

1-288

1-288

Plate Count

288

288

Empty Wells

N/A

N/A

Non-Recombinant Clones

8847 (8%)

8847 (8%)

Non-Insert Clones

N/A

N/A

Recombinant Clones

101745

101745

Average Insert Size

138 Kbp

136 Kbp

Genomic Coverage

12X

12X

 

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