Date 1" 24

Expt. No. Page No.

Aim the Eprinest i

Paparatien DH5a E coli comypetent Caly fot ttonslotnstion.
Praincile i- Competence is te ability Call to toke
extra celulor DNA. Iatzaducion exogengus DNA
inty botenia called tons fomoion: Comyetan e
Con he induced a ad h Cacla teeat ment
Cells becoma permeoble plosmid when ekposed
heat shock Selecton ttanslomd
Cels done he morker
in the plomid- A bocteriuy hating plesmid etn
Anpicillin Nesistonca Gon gw on anacitis Contoining
plates Ond thus selected .

Materia Reguireh i
’ cacl, (1M
’Souhon -I (o:jM Cal,)

Brocede i
1DHSa E.coi Calls iocudated in Yme LB madia
em glycorol stocd (3 tast tehes t 11 Conthol tube
Jhe celly lee 37c ot 220 pm
shoker
3. 5m himany Cultee Colls Were Sub Cutured en
250 LB mediu ond in Cubated a t 37"c uth

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Colcwatons :

oD Pinoy atre 5
OD for Seeondoy cutere s
Reqwud volun Secemdoy Gutu 250m

2= 5m
 Page No.
Expt. No,

QRo pmt the ODeoo O.6.

Shoking
3 The Cuttue Woo trons<ered in so md slerile foleen
ond od yo00 2pm for IS min
tube Centifuqd
4 7he boteial pellet wos Suspendecd 5m
Solution I uuith gente Suitling
5. he Cells for 5 minute
6he
he cells wete Spin -doun agoin at 4ooo tpm to
Ond
t The pelet sus pended ia I md
incubated on ie to 3o mins
Wes aigvated 1-5 m MCT ond the tubes
lwee frosen at -&o°c"

Rasults i
7lhe Competns peponed DHsA Gell eie be tested
isuyh ±nsjomaton ond furthet y ialating the
plosid ioterest DHSA Cels

Concusien i
ha phsa Conpetunt Cells
Epeinent geloted to
ta honyomation

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 Date

Expt. No. Page No.

mixing, Cols uith dieent ssehions should be Caiad

Qut iaside the Looingr ow heod
’ The MCTs Should be pre -cooled the
Competeat celo
The holessd Cella Should be kept co
|Experineot

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