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Genomic library

A enomic library is a population oI host bacteria, each oI which carries a DNA
molecule that was inserted into a cloning vector, (A c|on|ng vector ls a small plece
of unA lnLo whlch a forelgn unA fragmenL can be lnserLed 1he lnserLlon of Lhe
fragmenL lnLo Lhe clonlng vecLor ls carrled ouL by LreaLlng Lhe vehlcle and Lhe
forelgn unA wlLh a resLrlcLlon enzyme LhaL creaLes Lhe same overhang Lhen
llgaLlng Lhe fragmenLs LogeLher 1here are many Lypes of clonlng vecLors
CeneLlcally englneered plasmlds and bacLerlophages (such as phage ) are
perhaps mosL commonly used for Lhls purpose) such that the collection oI cloned
DNA molecules represents the entire genome oI the source organism. This term
also represents the collection oI all oI the vector molecules, each carrying a piece
oI the chromosomal DNA oI the organism, prior to the insertion oI these molecules
into the host cells.
%ypes of vectors
1. !lasmids: ( n mlcroblology and geneLlcs a |asm|d ls a unA molecule LhaL
ls separaLe from and can repllcaLe lndependenLly of Lhe chromosomal
unA 1hey are doublesLranded and ln many cases clrcular lasmlds
usually occur naLurally ln bacLerla ). Have a capacity oI 15 kb.
2. !hage (lambda)s: (A acter|oage ls any one of a number of vlruses LhaL
lnfecL bacLerla 8acLerlophages are among Lhe mosL common blologlcal
enLlLles on LarLh 1he Lerm ls commonly used ln lLs shorLened form f).
Have a capacity oI 25 kb.
3. osmids or Fosmids(A cosm|d flrsL descrlbed by Colllns and Pohn ln 1978
ls a Lype of hybrld plasmld (ofLen used as a clonlng vecLor) LhaL conLalns n
n unA sequences orlglnally from Lhe Lambda phage Cosmlds can
be used Lo bulld genomlc llbrarles). Have a capacity oI 35-45 kb.
4. Bacterial artiIicial chromosomes(A acter|a| art|f|c|a| cromosome (8AC) ls
a unA consLrucL based on a funcLlonal ferLlllLy plasmld (or lplasmld) used
for Lransformlng and clonlng ln bacLerla usually n) (BA): Have a
capacity oI 50-300 kb.
5. Yeast artiIicial chromosomes (A east art|f|c|a| cromosome (AC) ls a
vecLor used Lo clone unA fragmenLs larger Lhan 100 kb and up Lo 3000 kb


?ACs are useful for Lhe physlcal mapplng of complex genomes and for Lhe
clonlng of large genes)(YA): Have a capacity oI 300- ~1500 kb.
6. Human artiIicial chromosomes (A man art|f|c|a| cromosome (nAC) ls a
mlcrochromosome LhaL can acL as a new chromosome ln a populaLlon of
human cells 1haL ls lnsLead of 46 chromosomes Lhe cell could have 47
wlLh Lhe 47Lh belng very small roughly 610 megabases ln slze and able Lo
carry new genes lnLroduced by human researchers) (HA): Have a capacity
oI ~2000 kb.
reatin a library
The DNA molecules oI an organism oI interest are isolated. The DNA molecules
are then partially digested by an endonuclease restriction enzyme. Sometimes, the
DNA molecules are digested Ior diIIerent lengths oI time in order to ensure that all
the DNA has been digested to manageable sizes. The digested DNA molecules are
separated by size using agarose electrophoresis, and a suitable range oI lengths oI
DNA pieces are isolated and ligated into vectors. The vectors can then be taken up
by suitable hosts.The hosts are kept in liquid media and can be Irozen at -80 Ior
a long period oI time. Usually the hosts are bacteria that do not contain any
plasmids, so as to be sensitive to antibiotics.
The process oI subdividing genomic DNA into clonable elements and inserting
them into hosts is called creating a library, a clone bank or a gene bank. A
complete library oI host cells will contain all oI the genomic DNA oI the source
organism.