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 Home > Resources > Libraries
   VMRC-17: Rat (F) (Rattus norvegicus,FHH/Eur) BAC Library   

This rat (Rattus norvegicus) BAC library was constructed by Alicia Hill-Force and Joshua Danke in Chris Amemiya’s laboratory at the Benaroya Research Institute at Virginia Mason (BRI). This library was designated VMRC-17. A freshly dissected brain tissue sample was obtained from a male rat (strain FHH/EUR) from Drs. Joe Lazar and Howard Jacob (Medical College of Wisconsin). Nuclei were isolated using a Dounce homogenizer, embedded in Incert agarose; high molecular weight DNA was prepared in situ in the agarose. Generation of the library followed the general cloning approach developed in Pieter de Jong’s laboratory (Osoegawa et al., 1998). EcoRI fragments from the appropriate size fraction were cloned into the EcoRI site of the pCCBAC vector (Epicentre Technologies). The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 384-well microtiter dishes. Analysis of ~ 200 randomly selected clones from the library were used to assess the quality of the library. This work was funded by NIH grant HG02526 (Chris T. Amemiya, BRI).

Segment

1

2

All

Vector

pCC1BAC

pCC1BAC

pCC1BAC

Restriction Enzyme

EcoRI

EcoRI

EcoRI

DNA Source

brain

brain

brain

Plate Numbers

1-288

289-576

1-576

Plate Count

288

288

576

Empty Wells

0 (0%)

0 (0%)

0 (0%)

Non-Recombinant Clones

N/A

N/A

N/A

Non-Insert Clones

Approx. 7000 (6.33%)

Approx. 7000 (6.33%)

Approx. 14000 (6.33%)

Recombinant Clones

103592

103592

207184

Average Insert Size

148 Kbp

148 Kbp

148 Kbp

Genomic Coverage

5X

5X

10X

 

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