密歇根大学生物系实验室的常用试剂配方.docx

密歇根大学生物系实验室的常用试剂配方.docx

《密歇根大学生物系实验室的常用试剂配方.docx》由会员分享，可在线阅读，更多相关《密歇根大学生物系实验室的常用试剂配方.docx（6页珍藏版）》请在冰点文库上搜索。

密歇根大学生物系实验室的常用试剂配方

TableofContents

LBMedium.............................................................................................1

NZMedium............................................................................................2

SMBuffer................................................................................................3

SETBuffer...............................................................................................4

6XPrehybSoln.......................................................................................5

10XTBE...................................................................................................6

10XTAE..................................................................................................7

20XSSC...................................................................................................8

1%SDS,0.2MNaOH............................................................................9

14%PEG（8000）,2MNaCl,10mMMgSO4.....................................10

20%SDS.................................................................................................11

1.0MTris,pH8.0,1.5MNaCl...........................................................12

10mMTris-HCl,pH7.5,10mMMgSO4...........................................13

10mMTris,50mMEDTA,pH7.5....................................................14

10mMTris-HCl,1mMEDTA,pH7.5.............................................15

3MSodiumAcetate,pH4.8...............................................................16

Electrophoresisdye...............................................................................17

LabellingStopdye.................................................................................18

Sequencinggeldye...............................................................................19

5%Acrylamide......................................................................................20

6%AcrylamideinTBE,50%Urea.....................................................21

40%Acrylamide....................................................................................22

LBMedium（1Liter）

10gBacto-tryptone

5gBacto-yeastextract

10gNaCl

Forfortyplatesadd1%agar--1g.Autoclavemedia.Whencool,addampicillinandpourplates.For1Lofmedia,add1.8mLamp.

NZMedium（500mL）

5gBacto-tryptone

2.5gBacto-yeastextract

2.5gNaCl

1.25gMgSO4

For20platesadd1.2%agar--6g.Autoclaveandpourplatesat50oC

SMBuffer（1L）

5.8gNaCl

1.2gMgSo4

50mL1MTris-HCl,pH7.5

0.1gGelatin（doesn'tdissolve）

Autoclave

Usedforphagedilutionandstorage.

SETBuffer

50mMTris-HCl,pH8.0,50mMEDTA,20%w/vSucrose

tomake200mL:

40gSucrose

10mLof1MTris

20mLof0.5MEDTA,disodiumsalt

bringto200mLwithH20

6XPrehybridizationSolution

tomake500mL

300mLddH20

150mL20XSSC

50mL50XDenhardt'ssolution

1mL0.5MEDTA（disodiumsalt）

2.5mL20%SDS

6XreferstotheconcentrationofSSC

10XTBEBuffer（forpolyacrylamidegels）

tomakeoneliter:

60.75gTris

3.7gEDTA（tetrasodiumsalt）

30gBoricacid

10XTAEBuffer（Foragarosegels）

tomakeoneliter:

48.20gTris

6.75gNaAce

3.75gEDTA（disodiumsalt）

AdjustpHto7.6withaceticacid.（Approx.20mL）

20XSSC

tomakeoneliter:

175.3gNaCl

88.2gNaCitrate

addwatertobringvolumetooneliter.

adjusttopH7.0withHCl.

1%SDS,0.2MNaOH

tomake100mL:

93mLddH20

5mL20%SDS

2mL10MNaOH

14%PEG（8000）,2MNaCl,10mMMgSO4

tomakeoneliter:

140gPEG

117gNaCl

2.46gMgSO4

ForuseinphageDNApreparation.

20%SDS

tomale250mL:

50gofSDSinabeaker

AddstirbarandH20last.

ThissolutionwillhavetobeheatedfortheSDStodissolve.

1.0MTris,pH8.0,1.5MNaCl

tomakeoneliter:

121.1gTrizma

87.6gNaCl

inavolumeofwaterlessthan1L.AdjustpHwithHCl,thenbringto1LwithH20

10mMTris-HCl,pH7.5,10mMMgSO4

tomakeoneliter:

10mL1MTris-HCl

2.46gMgSO4

foruseinphageDNApreparation

10mMTris,50mMEDTA,pH7.5

tomake200mL:

2mL1MTris

20mL0.5MEDTA（tetrasodiumsalt）

178mLddH20

adjustpHwithHCl.

10mMTris-HCl,1mMEDTA,pH7.5

tomake200mL:

2.0mL1MTris-HCl,pH7.5

0.4mL0.5MEDTA

197.6mLddH20

3MSodiumAcetate,pH4.8

tomakeoneliter:

408.1gNaAce（trihydrate;getscoldinsoln）

about700mLH20

adjustpHwithglacialaceticacid（takesalot）

MeasuretrupHbydilutionwithwater;rangewillbebetween4.8and5.5.

ElectrophoresisDye

tomake4mL:

3mL50mMEDTA,10mMTris-HCl,pH8.0

1mLglycerol

20LBPB

10LXylenecyanol

Stopdyeforlabelledprobe

1mL50mMEDTA,10mMTris,pH7.5-8.5

about200lglycerol

addafewgrainsofbluedextran（8000）

Sequencinggeldye

forapprox1mL:

1mLformamide

10Lxylenecyanol

10lBPB

3L10MNaOH

5%acrylamide

tomake200mL:

20mL10XTBE

25mL40%acrylamide

155mLH20

6%AcrylamideinTBE,50%Urea

tomake500mL:

50mL10XTBE

75mL40%acrylamide

250gUrea

bringto500mLwithH2O

40%Acrylamide（38:

2acrylamide:

bisacrylamide）

tomake200mL:

76gacrylamide

4gbisacrylamide

bringto200mLwithH2O