autoclave是什么意思oclave在线翻译读音

ElectrocompetentCellPreparation

outbacterialstraintobeusedonLMplates(notLB).Restreak(without

cooling)asinglecolonyagainonLMthenextdaytofurtherincreasecompetency.

2。PrepareseverallitersofddH

2

0，and~100mlof10%glycerolaheadoftimeand

storeincoldroomthedaybeforemakingcompetentcells。Donotautoclave

glycerol，filterthrough0。45mfilter。.

10fresh，juicy,activelygrowingbacterialcoloniesan望驿台元稹 dculturethemin100

mlpre-warmedSOBin1000mlflask，shakingvigorouslyat37C(250rpmOK）.

Thistake古诗词摘抄及赏析大全 s1—3hours.（Seealternatestep3Abelow）。

lturebeginningtogetc诡谲怎么读 loudy(butO。D。

600

<0。25)，splitintofour

250mlculturesofprewarmedSOB,forfinalvolumeof1。0liter。Usefour2.0liter

flasks。Continuetoshakevigorouslyat37icallycheckO。D。

600

.(See

alternatestep4Abelow）。

5。WhenO.D.

600

0.50-0。60，lltake1—3morehours.

CollectintochilledGS—mall

subsequentstepsat4Coroniceuntilfreezingofcells.

4,000rpmat4rainswillpelletatthisspeed—but

somedon’cloudy,try5,000rpm。Notethatasthecellsbecomemore

thoroughlywashed,higherspeedspinsarenecessary.

7摊破浣溪沙拼音 。Resuspendpelletsin500mlcoldddH

2

againinGS—3at5，000rpm，

4C,20minutes.

8。Resuspendpelletsin500mlcoldddH

2

againinGS—3at6,000rpm,

4C，20minutes.

9。Resuspendpelletsin25ml10%glycerolinddH

2

0。Filterglycerolsolution

through0。45mfilterbeforeusing。Donotautoclaveglycerol。Use50ml

disposablecentrifugetubesandspin6，500rpm，at4C,25minutes，inHS-4

tubeforcellsandabalance.

endcellpelletin3。0mlssterile10%200l

aliquotsintomicrofugetubes，bespre—labelledwith

strainname，andnotebookpagenumber。

irectlyin-80Cfreezer。

AlternateApproachwithOvernightCulture：Substitutesteps3A—4Aforsteps3-4

above.

efreshcolonyfromLMplate（nevercooled）,cultureovernightin100

mlpre—warmedSOBin1000mlflask，shakingvigorouslyat37C(250rpmOK).

Startthisovernightcultureaslat意思相反的词语 easpossiblebeforegoinghome.

tday，firstthinginthe描写庐山的古诗 A.M。,inoclulatefour250mlculturesof

pre—r2。0

literflasks。Shakevigorouslyat37C（250rp咏月的诗句古诗 mOK）。PeriodicallycheckO。

D。

600

.lturestepwilltake3-5hours。

LBMEDIUM

for1Liter

=10gbacto-tryptone

5gbacto-yea己亥杂诗其五拼音版 stextract

10gNaCl

AdjustpHto7。0with5NNaOH（about0.2ml翻箱倒柜是什么意思 )。Autoclave。

LabdistilledwaterOK

LMPlates

Thesplatesareforculturingbacteriatobeusedformakingcompetentcellsor

electrocompetentcells.

for1liter

=10gbacto—tryptone

5gyeastextract

0.58gNaCl

15gBactoAgar

Add800mltapH

2

0

pHto7。0

Adjustto1。0literwithtapH

2

0

Autoclave

Aft道字组词 erautoclaving，whencooling,add10mls1。0MMgSO

4

Note—theseplatesdon\'tworkwithtetracyclinesincetheycontainmagnesium.

SOBMEDIUM

for1liter

=20gbacto—tryptone

5gbacto—yeastextract

0。5gNaCl

Disolvein950mlwater

Make250mMKCl：

1.86gdisolvedin100mlwater

Make5NNaOH：

2gfor10ml

Make2MMgCl2：

ave。

Add10mlof250mMKCl

AdjustpHto7。0with5NNaOH（about0。2ml)

Bringvolumeupto1liter

Autoclave

SOCMEDIUM

for1liter

ThesameasSOBmediumwiththeadditionof20mMglucose。

Make1Mglucose:

18gin90mlofwater

Bringvolumeto100ml

Filterthrough0。22mfilter

AutoclaveSOBmedium

Whencooledto60Corless，add20mlsterileglucoseperliter

Remembertoadd5mlsterileMgCl2justbeforeuse.