Erman's Lab:DNA Miniprep with Alkaline Lysis - Revision history

Vaishnavi Ananth: /* BioCoder version */

2009-11-19T09:52:28Z

BioCoder version

←Older revision		Revision as of 09:52, 19 November 2009
Line 115:		Line 115:
	#Please refer to Maniatis' Molecular Clonning: A Labratory Manual for further information		#Please refer to Maniatis' Molecular Clonning: A Labratory Manual for further information

-	==~~BioStream~~ version==	+	==BioCoder version==
-	Following is the Erman's Lab:DNA Miniprep with Alkaline Lysis protocol in ~~BioStream~~, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in ~~BioStream~~ (see Source code). More information about ~~BioStream~~ can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]	+	Following is the Erman's Lab:DNA Miniprep with Alkaline Lysis protocol in BioCoder, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioCoder (see Source code). More information about BioCoder can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
	====Text Output====		====Text Output====
	[[Erman's Lab:DNA Miniprep with Alkaline Lysis protocol]]		[[Erman's Lab:DNA Miniprep with Alkaline Lysis protocol]]

Vaishnavi Ananth at 05:56, 20 October 2009

2009-10-20T05:56:06Z

←Older revision		Revision as of 05:56, 20 October 2009
Line 114:		Line 114:
	==References==		==References==
	#Please refer to Maniatis' Molecular Clonning: A Labratory Manual for further information		#Please refer to Maniatis' Molecular Clonning: A Labratory Manual for further information
		+
		+	==BioStream version==
		+	Following is the Erman's Lab:DNA Miniprep with Alkaline Lysis protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.[[User:Vaishnavi Ananth\|Vaishnavi]]
		+	====Text Output====
		+	[[Erman's Lab:DNA Miniprep with Alkaline Lysis protocol]]
		+	====Source Code====
		+	[[Erman's Lab:DNA Miniprep with Alkaline Lysis protocol - source code]]

Emre Deniz at 19:45, 17 October 2009

2009-10-17T19:45:49Z

←Older revision		Revision as of 19:45, 17 October 2009
Line 4:		Line 4:

	==Materials==		==Materials==
-	*STE Stock From Stock(for 50ml) Final	+	*STE
-	~~TrisCl~~ 1M ~~TrisCl~~ 0.5ml 10mM	+	{\|
-	~~NaCl~~ 5M NaCl 1ml 100mM	+	\|-
-	~~EDTA~~ 0.5M EDTA 0.1ml 1mM	+	! Stock
		+	! From Stock(for 50ml)
		+	! Final
		+	\|-
		+	\| 1M Tris(pH8)
		+	\| 0.5ml
		+	\| 10mM
		+	\|-
		+	\| 5M NaCl
		+	\| 1ml
		+	\| 100mM
		+	\|-
		+	\| 0.5M EDTA
		+	\| 0.1ml
		+	\| 1mM
		+	\|}

-	*ALKALINE LYSIS1 Stock From Stock(for 50ml) Final
-	~~TrisCl 1M Tris(pH8) 1ml 20mM~~
-	~~EDTA 0.5M EDTA 1ml 10mM~~
-	~~Glucose FW=108.2g 0.45g 50mM~~

-	*ALKALINE LYSIS 2 Stock From Stock(for ~~10ml~~) Final	+	*ALKALINE LYSIS 1
-	~~NaOH 5N NaoH~~ 0.~~4ml~~ 0.2N	+	{\|
-	~~SDS 10%SDS~~ 1ml 1%	+	\|-
-	~~dH2O Up to 10ml~~	+	! Stock
		+	! From Stock(for 50ml)
		+	! Final
		+	\|-
		+	\| 1M Tris(pH8)
		+	\| 1ml
		+	\| 20mM
		+	\|-
		+	\| Glucose FW:108.2
		+	\| 0.45g
		+	\| 50mM
		+	\|-
		+	\| 0.5M EDTA
		+	\| 0.1ml
		+	\| 10mM
		+	\|}

-	*ALKALINE LYSIS 3 Stock From Stock(for 50ml) Final
-	~~KAc 5M KAc 30ml 3M~~
-	~~Acetic acid Glacial ac ac 5.75ml 5M acetate~~
-	~~dH2O Up to 50ml~~

		+	*ALKALINE LYSIS 2
		+	{\|
		+	\|-
		+	! Stock
		+	! From Stock(for 10ml)
		+	! Final
		+	\|-
		+	\| 5N NaOH
		+	\| 0.4ml
		+	\| 0.2N
		+	\|-
		+	\| 10% SDS
		+	\| 1ml
		+	\| 1%
		+	\|-
		+	\| dH<sub>2</sub>O
		+	\| up to 10ml
		+	\|
		+	\|}
		+
		+
		+	*ALKALINE LYSIS 3
		+	{\|
		+	\|-
		+	! Stock
		+	! From Stock(for 50ml)
		+	! Final
		+	\|-
		+	\| 5M KAc
		+	\| 30ml
		+	\| 3M
		+	\|-
		+	\| Glacial ac ac
		+	\| 5.75ml
		+	\| 5M acetate
		+	\|-
		+	\| dH<sub>2</sub>O
		+	\| up to 50ml
		+	\|
		+	\|}

	=='''Procedure'''==		=='''Procedure'''==

Emre Deniz: New page: ==Overview== Miniprep DNA from E. Coli ==Materials== STE Stock From Stock(for 50ml) Final TrisCl 1M TrisCl 0.5ml 10mM NaCl 5M NaCl 1ml 100mM EDTA 0.5M EDTA 0.1ml 1mM ALKALINE LYSIS1 S...

2009-10-17T19:00:39Z

New page: ==Overview== Miniprep DNA from E. Coli ==Materials== *STE Stock From Stock(for 50ml) Final TrisCl 1M TrisCl 0.5ml 10mM NaCl 5M NaCl 1ml 100mM EDTA 0.5M EDTA 0.1ml 1mM *ALKALINE LYSIS1 S...

New page

==Overview==

Miniprep DNA from E. Coli

==Materials==
*STE Stock From Stock(for 50ml) Final
TrisCl 1M TrisCl 0.5ml 10mM
NaCl 5M NaCl 1ml 100mM
EDTA 0.5M EDTA 0.1ml 1mM

*ALKALINE LYSIS1 Stock From Stock(for 50ml) Final
TrisCl 1M Tris(pH8) 1ml 20mM
EDTA 0.5M EDTA 1ml 10mM
Glucose FW=108.2g 0.45g 50mM

*ALKALINE LYSIS 2 Stock From Stock(for 10ml) Final
NaOH 5N NaoH 0.4ml 0.2N
SDS 10%SDS 1ml 1%
dH2O Up to 10ml

*ALKALINE LYSIS 3 Stock From Stock(for 50ml) Final
KAc 5M KAc 30ml 3M
Acetic acid Glacial ac ac 5.75ml 5M acetate
dH2O Up to 50ml

=='''Procedure'''==
==Isolation of Mononuclear Cells==

#o/n grow single colony in 2ml of LB (+ antibiotics)
#1.5 ml into eppendorf.
#Pellet cells at max speed in cold room for 1.5 min.
#Discard supernatant , leave pellet as dry as possible.
#Resuspend pellet in 100 μL Alkaline Lysis SLN1,vortex.
#Add 200μL freshly prepared AL2, mix by inverting 5-6 times(DO NOT vortex!)
#Put on ice.
#Add 300μL AL3,invert tubes to mix.
#Incubate on ice for 5 min.
#Centrifuge at max speed for 5 min in cold room.
#Take the supernatant into a new tube.
#Add 900 isopropanol at RT. Vortex
#Centrifuge at max speed for 10 min at RT.
#Rinse the pellet with 1ml 70%EtOH.
#Centrifuge at max speed for 5 min at RT.
#Air dry the pellet.
#Dissolve each in 50μL TE OR H<sub>2</sub>O.

==Notes==
#Store solutions at 4 degrees and each time freshly prepare the Buffer 2.

==References==
#Please refer to Maniatis' Molecular Clonning: A Labratory Manual for further information

[[Category:Protocol]]

[[Category:Miniprep]]

[[Category:Maniatis]]

[[Category:DNA]]

[[Category:Escherichia coli]]

Erman's Lab:DNA Miniprep with Alkaline Lysis - Revision history

Vaishnavi Ananth: /* BioCoder version */

Vaishnavi Ananth at 05:56, 20 October 2009

Emre Deniz at 19:45, 17 October 2009

Emre Deniz: New page: ==Overview== Miniprep DNA from E. Coli ==Materials== *STE Stock From Stock(for 50ml) Final TrisCl 1M TrisCl 0.5ml 10mM NaCl 5M NaCl 1ml 100mM EDTA 0.5M EDTA 0.1ml 1mM *ALKALINE LYSIS1 S...

Emre Deniz: New page: ==Overview== Miniprep DNA from E. Coli ==Materials== STE Stock From Stock(for 50ml) Final TrisCl 1M TrisCl 0.5ml 10mM NaCl 5M NaCl 1ml 100mM EDTA 0.5M EDTA 0.1ml 1mM ALKALINE LYSIS1 S...