biology report -group 1

8/20/2019 Biology Report -Group 1

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BIOLOGY REPORT

Experiment date: 3-11-2015 Group: 1Fu name: !"u#$n %&n' (an" )*+n" !", T.n !"u#$n T'an' T/n"

I:2013303I:201335I:20133

%a: B4E %56

I7 8ed oution:1. LB broth medium (to culture E.coli):

+ Peptone/tryptone: 1 g

+ Yeast extract : .! g

+ "a#l : 1 g

$issol%e all components in & ml '* then ill up to 1 ml. ,utoclare at 11o#

or - min.

. Lysis buer:+ m 0ris'#l* p' &.

+ 2 m 3$0,* p' &.

+ 1! m "a#l

+ 14 5$5

-. ther solutions: ! #'-#6* p' .&7 8sopropanol 147 3thanol 94

. P# reagents: (master mix)

+ ! m d"0Ps

+ ! µ Primers

+ 1; Buer + 0a< $", polymerase

!. 3lectrophoresis buer: 1; 0,3

+ 0risbase : .& g

+ =lacial ,cetic ,cid : 1.> g

+ 3$0, : .> g


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$issol%e and ill up by 1 l o '

2. =ram stain solutions

II7 Pratie 1: O9eration o; 9ateria 9# miroope

• Procedure:

?'eatix@ the slide Aith the specimen by passing it o%er a heat source* such as a

lame* se%eral times using a orceps. 0he slide should be passed %ery


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'igh density

0hin cellAall ⇒ ater Aashing by alcohol* the stain is remo%ed

+,t the end o the gram staining procedure* gram negati%e cells Aill be stained

a reddishpin color.

,nd here are the pictures o gramnegati%e* that Ae too in the laboratory:

=ram G negati%e microorganisms ( E.coli)


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=ram G positi%e microorganisms (possibly ungi) stained deep %iolet to blue

#omments:

He got something ailed here* may be in the staining process* so the gram

negati%e is mixed Aith the grampositi%e. 8t lead to the bad results lie thecolor o the bacteria (8t is pin* and a little bit o blue* not as the real color o

the bacteria).,nd in the staining process* the decoloEier is let too long. 8 the

decoloriEer is let on too long* e%en gram positi%e cells Aill lose the crystal

%iolet and Aill stain red. But Ae still get some comments:

+od orm I Lshaped orm

+5ome indi%iduals are deorm and old

+0hicer cellAall

+$ecoloriEing the cell causes this thic cell Aall to dehydrate and shrin*Ahich closes the pores in the cell Aall and pre%ents the stain rom exiting the

cell

+LoA density and scattering arrangement

=ram G positi%e microorganisms (possibly ungi)

III7 Pratie 2: !< extration ;rom 9ateriaProcedure:


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'ar%est the E.coli cell pellets rom o%ernight culture by centriugation at ! rpm

or ! min

$issol%e the pellet in ! Jl lysis buer. 0he tube is then let at room temperature

or 1 minutes

,dd 1! Jl #'-#6* p' K .& and mix lightly ,dd 2! Jl #8 and mix Aell by hand in%ersion

5pin the tube in the centriuge at 1 rpm or ! minutes

#ollect and transer the supernatant to another 1.!ml 3ppendor tube

,dd an e


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Reut: =roup 1: !9&.& .- .!

+,t $", sample: the %alues o 2/&* 2/- are at accepted %alues

(.-N1.& and .!N.) KN $", is pure.

+ 0he 2/- %alues or ?pure@ nucleic acid are oten higher than the respecti%e

2/& %alues. 3xpected 2/- %alues are commonly in the range o ... 8

the ratio is appreciably loAer than expected* it may indicate the presence o

contaminants Ahich absorb at - nm.

P%R:


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#oncentration 2/& 2/-

E.coli !92.- 1.2! 1.>


8/11

)7 Pratie 5:


9/11

P# cycling

17 > o#: ! minutes27 > o#: 1 minute37 21 o#: 1 minute7 9 o#: 1 minute* repeat rom step or -! times57 9 o#: 9 minutes

Reut:

5tructure o Aells on electrophoresis image

=roup 1 =roup =roup -0est

arer +

Hells 1 - ! 2 9 & >

P# $"

,P# $", P# $", P# $",


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• O9eration: agarose gel electrophoresisBasically* Ae can state that the tAo experiments o - groups Aere successul

E.coli $",:

Because o some reasons ($", Aas dried or too long or Aell breaing* etc.)*

$", image %ia electrophoresis is not distributed as expected. $", parts stic

together and unable to mo%e along the electric ield.

- groups has the same type o image7 hoAe%er* group - has the best result.


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ur group result is

biology report -group 1

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