Hackteria - Recent changes [en]

DIY Fermentation or the Art of making Wine

2010-09-06T16:24:29Z

← Older revision		Revision as of 16:24, 6 September 2010
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-	[[File:mach_deinen_wein_doch_selbst.jpg]]	+	[[File:mach_deinen_wein_doch_selbst.jpg\|1024px]]

GFP Protocol Jugaad

2010-09-06T09:12:24Z

← Older revision		Revision as of 09:12, 6 September 2010
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	[[File:MG_7190.JPG]] [[File:MG_7186.JPG]]		[[File:MG_7190.JPG]] [[File:MG_7186.JPG]]

		+	[[File:DSC02301.JPG]]


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	10. When using the water bath run it till it reaches the required temp. and then use it.		10. When using the water bath run it till it reaches the required temp. and then use it.
-	~~[[File:Example.jpg]]~~

File:DSC02301.JPG

2010-09-06T09:12:17Z

uploaded "[[File:DSC02301.JPG]]"

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DIY Sterlisation Hood

2010-09-06T08:17:45Z

STERLISATION HOOD:

← Older revision	Revision as of 08:17, 6 September 2010
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	+	Trust me this sounds easy but you need alot of patience and material to do this.

GFP Protocol Jugaad

2010-09-06T08:03:33Z

← Older revision		Revision as of 08:03, 6 September 2010
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	[[File:MG_7020.JPG]] [[File:MG_7028.JPG]]		[[File:MG_7020.JPG]] [[File:MG_7028.JPG]]

-	. [[File:MG_7052.JPG]]	+	[[File:DSC02293.JPG]] [[File:MG_7052.JPG]]

	[[File:MG_7075.JPG‎]] [[File:MG_7083.JPG]]		[[File:MG_7075.JPG‎]] [[File:MG_7083.JPG]]

-	. [[File:MG_7052.JPG]]	+	[[File:DSC02295.JPG]] [[File:MG_7052.JPG]]

	[[File:MG_7174.JPG]]		[[File:MG_7174.JPG]]
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	[[File:MG_7176.JPG]] [[File:MG_7177.JPG]]		[[File:MG_7176.JPG]] [[File:MG_7177.JPG]]

-	. [[File:MG_7174.JPG]]	+	[[File:DSC02296.JPG]] [[File:MG_7174.JPG]]

	[[File:MG_7163.JPG]] [[File:MG_7028.JPG]]		[[File:MG_7163.JPG]] [[File:MG_7028.JPG]]
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	[[File:MG_7190.JPG]] [[File:MG_7186.JPG]]		[[File:MG_7190.JPG]] [[File:MG_7186.JPG]]

-	~~.(incubate pic)~~


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	10. When using the water bath run it till it reaches the required temp. and then use it.		10. When using the water bath run it till it reaches the required temp. and then use it.
		+	[[File:Example.jpg]]

File:DSC02296.JPG

2010-09-06T07:45:43Z

uploaded "[[File:DSC02296.JPG]]"

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GFP Protocol Jugaad

2010-09-06T06:32:59Z

← Older revision		Revision as of 06:32, 6 September 2010
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	[[File:MG_7213.JPG]] [[File:MG_7221.JPG]]		[[File:MG_7213.JPG]] [[File:MG_7221.JPG]]

-	[[File:MG_7225.JPG]] ~~pippet out on plate~~	+	[[File:MG_7225.JPG]] [[File:3.JPG]]
		+
		+	[[File:4.JPG]] [[File:5.JPG]]
		+
		+	[[File:6.JPG]] [[File:7.JPG]]

-	~~spreader incubate~~


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	== '''Precautions:''' ==		== '''Precautions:''' ==

-	1. ~~work~~ in a closed room with al the equipment in it. Don't run around for things.	+	1. Work in a closed room with all the equipment in it. Don't run around for things.

-	2. ~~keep~~ the room clean. No eatables, foot ~~ware~~ allowed.	+	2. Keep the room clean. No eatables, foot wear allowed.

-	3. ~~run~~ UV light for about half an hour before starting experiment.	+	3. Run UV light for about half an hour before starting experiment.

	'''Note:''' do not stand close to the UV when it is on. use a uv filter to cover all sides of the box and if posible cover the incubator and walls also.		'''Note:''' do not stand close to the UV when it is on. use a uv filter to cover all sides of the box and if posible cover the incubator and walls also.
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	4. Clean hands with alcohol before starting anything.		4. Clean hands with alcohol before starting anything.

-	5. ~~before~~ using glassware autoclave them (wrap in paper and boil in cooker for 3 wistles.)	+	5. Before using glassware autoclave them (wrap in paper and boil in cooker for 3 wistles.)
		+
		+	6. All work is done in the hood to avoid external contamination.

-	6. ~~all work is done~~ in ~~the hood to avoid external contamination.~~	+	7. Sterilize everything in alcahol and flame before use

-	7. ~~keep~~ someone beside you to give and take things from you while working in the hood. Don't keep moving your hands in and out of the hood.	+	8. Keep someone beside you to give and take things from you while working in the hood. Don't keep moving your hands in and out of the hood.

-	8. ~~keep~~ hood closed when not in use	+	9. Keep hood closed when not in use

-	9. ~~when~~ using the water bath run it till it reaches the required temp. and then use it.	+	10. When using the water bath run it till it reaches the required temp. and then use it.

File:7.JPG

2010-09-06T06:32:39Z

uploaded "[[File:7.JPG]]"

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Main Page

2010-09-06T03:53:09Z

← Older revision		Revision as of 03:53, 6 September 2010
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	\|-		\|-
	\| '''Project 19: [[DIY Water Bath]]'''		\| '''Project 19: [[DIY Water Bath]]'''
-	[[File:~~DSC04036~~.JPG‎\|120px\|left]]	+	[[File:DSC04040.JPG‎\|120px\|left]]


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	\| '''Project 20: [[GFP Protocol Jugaad]]'''		\| '''Project 20: [[GFP Protocol Jugaad]]'''
	[[File:MG_7283.JPG‎\|120px\|left]]		[[File:MG_7283.JPG‎\|120px\|left]]
-		+	Home made GFP bacteria

	</div>		</div>
	<!-- Projects end -->		<!-- Projects end -->

GFP Protocol Jugaad

2010-09-04T08:52:38Z

← Older revision		Revision as of 08:52, 4 September 2010
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	'''Note:''' If observed add 254 nm of less intensity of glow		'''Note:''' If observed add 254 nm of less intensity of glow
	could vary.		could vary.
		+
		+	'''Note:''' The 3 controls:
		+
		+	'''Positive control:''' (Competent cells + Control DNA.)
		+
		+	-If there are no colonies it means there is something wrong with the cells.
		+
		+	'''Negative control:''' (only Competent cells.)
		+
		+	-If there are no colonies it means the cells are contaminated.
		+
		+	'''Double Negative control:''' (Empty LB-Amp plate.)
		+
		+	-If there is growth then there is external contamination.
		+
		+
		+	[[File:MG_7281.JPG]] [[File:MG_7269.JPG]]
		+
		+	[[File:MG_7282.JPG]]
		+
		+
		+
		+
		+
		+
		+
		+	== '''Precautions:''' ==
		+
		+	1. work in a closed room with al the equipment in it. Don't run around for things.
		+
		+	2. keep the room clean. No eatables, foot ware allowed.
		+
		+	3. run UV light for about half an hour before starting experiment.
		+
		+	'''Note:''' do not stand close to the UV when it is on. use a uv filter to cover all sides of the box and if posible cover the incubator and walls also.
		+
		+	4. Clean hands with alcohol before starting anything.
		+
		+	5. before using glassware autoclave them (wrap in paper and boil in cooker for 3 wistles.)
		+
		+	6. all work is done in the hood to avoid external contamination.
		+
		+	7. keep someone beside you to give and take things from you while working in the hood. Don't keep moving your hands in and out of the hood.
		+
		+	8. keep hood closed when not in use
		+
		+	9. when using the water bath run it till it reaches the required temp. and then use it.

File:MG 7282.JPG

2010-09-04T08:25:48Z

uploaded "[[File:MG 7282.JPG]]"

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GFP Protocol Jugaad

2010-09-04T08:10:39Z

← Older revision		Revision as of 08:10, 4 September 2010
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	. [[File:MG_7174.JPG]]		. [[File:MG_7174.JPG]]

-	[[File:MG_7028.JPG]]	+	[[File:MG_7163.JPG]] [[File:MG_7028.JPG]]
		+
		+


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	'''Transformation:'''		'''Transformation:'''

-	'''	+
-	16.''' Take a 2 vials of ligated sample and control DNA and 1 vial of only control DNA( positive control).	+	'''16.''' Take a 2 vials of ligated sample and control DNA and 1 vial of only control DNA( positive control).
	To this add 200 μl each of competent cells. Tap the		To this add 200 μl each of competent cells. Tap the
	vials gently and incubate on ice for 20 minutes.		vials gently and incubate on ice for 20 minutes.
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	bacteria to recover and express the protein.		bacteria to recover and express the protein.

-	'''21.''' Pipette 200 μl each from the vials transformed with the	+	'''Note:''' While the heat shock treatment is going on, prepare the LB-Amp agar(200 μl in 200 ml of LB agar.)
		+
		+	[[File:MG_7064.JPG]] [[File:MG_7133.JPG]]
		+
		+	[[File:MG_7142.JPG]] [[File:MG_7143.JPG]]
		+
		+	[[File:MG_7174.JPG]] [[File:MG_7175.JPG]]
		+
		+	[[File:MG_7177.JPG]] [[File:MG_7174.JPG]]
		+
		+	[[File:MG_7185.JPG]] [[File:MG_7191.JPG]]
		+
		+	[[File:MG_7190.JPG]] [[File:MG_7186.JPG]]
		+
		+	.(incubate pic)
		+
		+
		+
		+	'''21.''' liquify the already made LB-amp agar and pour about 20 to 25 ml of i in each plate.
		+
		+	'''Note:''' Wait for agar to solidify again.
		+
		+	'''22.''' Pipette 200 μl each from the vials transformed with the
	ligated mix onto LB-Amp plates and spread thoroughly		ligated mix onto LB-Amp plates and spread thoroughly
	using spreader/pipette.		using spreader/pipette.

-	'''22.''' Pipette 200 μl of LB onto a LB -Amp plate and spread	+	'''23.''' Pipette 200 μl of LB onto a LB -Amp plate and spread
	20 ìl of the cells transformed with control DNA. Label		20 ìl of the cells transformed with control DNA. Label
	this as positive control plate.		this as positive control plate.

-	'''23.''' Plate 200 μl of non-transformed cells onto another	+	'''24.''' Plate 200 μl of non-transformed cells onto another
	LB-Amp plate to check for contamination. Label this		LB-Amp plate to check for contamination. Label this
	as non-transformed (negative control) plate.		as non-transformed (negative control) plate.

-	'''24.''' Keep one plate with only LB-Amp Agar and nothing else. Name this as double negative control.	+	'''25.''' Keep one plate with only LB-Amp Agar and nothing else. Name this as double negative control.
		+
		+	'''26.''' Incubate the plates at 37°C, overnight.
		+
		+
		+	[[File:MG_7204.JPG]] [[File:MG_7206.JPG]]
		+
		+	[[File:MG_7213.JPG]] [[File:MG_7221.JPG]]
		+
		+	[[File:MG_7225.JPG]] pippet out on plate

-	~~'''25.''' Incubate the plates at 37°C, overnight.~~	+	spreader incubate


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	'''Moment of Truth'''		'''Moment of Truth'''

-	'''26.''' Observe the plates under UV-light (312 nm).	+	'''27.''' Observe the plates under UV-light (312 nm).

	'''Note:''' If observed add 254 nm of less intensity of glow		'''Note:''' If observed add 254 nm of less intensity of glow
	could vary.		could vary.

File:MG 7225.JPG

2010-09-04T08:01:08Z

uploaded "[[File:MG 7225.JPG]]"

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GFP Protocol Jugaad

2010-09-04T06:42:33Z

← Older revision		Revision as of 06:42, 4 September 2010
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	'''7.''' Chill the culture tubes on ice for 10-20 minutes.		'''7.''' Chill the culture tubes on ice for 10-20 minutes.

-	'''8.''' Transfer the culture aseptically into sterile vials, 5 vials with 5ml ~~and 5 vial with 10ml~~	+	'''8.''' Transfer the culture aseptically into sterile vials. (5 vials with 0.5ml each)

	'''9.''' Spin down at 6000 rpm for 8 minutes,		'''9.''' Spin down at 6000 rpm for 8 minutes,
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	33 ml of solution A, resuspend gently.		33 ml of solution A, resuspend gently.

-	'''12.''' Keep the ~~centrifuge tubes~~ on ice for 5 minutes.	+	'''12.''' Keep the vials on ice for 5 minutes.
-	Centrifuge at 8000 rpm for 15 minutes at 4°C or spin at	+	Centrifuge at 8000 rpm for 5 minutes at 4°C or spin at
	RT.		RT.

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	'''Note:''' Resuspension is to be done gently as cells		'''Note:''' Resuspension is to be done gently as cells
	are very fragile at this stage.		are very fragile at this stage.
		+
		+	[[File:MG_7020.JPG]] [[File:MG_7028.JPG]]
		+
		+	. [[File:MG_7052.JPG]]
		+
		+	[[File:MG_7075.JPG‎]] [[File:MG_7083.JPG]]
		+
		+	. [[File:MG_7052.JPG]]
		+
		+	[[File:MG_7174.JPG]]
		+
		+

	'''14.''' Heat inactivate the ligated samples at 65°C for		'''14.''' Heat inactivate the ligated samples at 65°C for
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	the vials on ice.		the vials on ice.

-	'''15.''' Aliquot 2 μl (10 ng) of control DNA each into 5 ~~prechilled~~	+	'''15.''' Aliquot 2 μl (10 ng) of control DNA each into 5 ligation vials
-	1.~~5 ml vials~~.	+
		+	[[File:MG_7176.JPG]] [[File:MG_7177.JPG]]
		+
		+	. [[File:MG_7174.JPG]]
		+
		+	[[File:MG_7028.JPG]]
		+


	'''Transformation:'''		'''Transformation:'''

-	~~'''Note:''' Use single aliquot of competent cells~~
-	~~(0.6 ml) for one set of transformation experiment.~~
	'''		'''
-	16.''' Take a ~~single vial~~ of ligated sample and control DNA.	+	16.''' Take a 2 vials of ligated sample and control DNA and 1 vial of only control DNA( positive control).
	To this add 200 μl each of competent cells. Tap the		To this add 200 μl each of competent cells. Tap the
	vials gently and incubate on ice for 20 minutes.		vials gently and incubate on ice for 20 minutes.
-	~~12 13~~

-	'''17.''' Label the remaining 200 μl of competent cells as ~~nontransformed~~	+
		+	'''17.''' Label the remaining 200 μl of competent cells as non transformed (negative control)
	cells, place on ice till the plating step (step		cells, place on ice till the plating step (step
-	24).	+	23).

	'''18.''' Tap all the vials gently and incubate on ice for		'''18.''' Tap all the vials gently and incubate on ice for
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	'''22.''' Pipette 200 μl of LB onto a LB -Amp plate and spread		'''22.''' Pipette 200 μl of LB onto a LB -Amp plate and spread
	20 ìl of the cells transformed with control DNA. Label		20 ìl of the cells transformed with control DNA. Label
-	this as control plate.	+	this as positive control plate.

	'''23.''' Plate 200 μl of non-transformed cells onto another		'''23.''' Plate 200 μl of non-transformed cells onto another
	LB-Amp plate to check for contamination. Label this		LB-Amp plate to check for contamination. Label this
-	as non-transformed plate.	+	as non-transformed (negative control) plate.

-	'''24.''' Incubate the plates at 37°C, overnight.	+	'''24.''' Keep one plate with only LB-Amp Agar and nothing else. Name this as double negative control.
		+
		+	'''25.''' Incubate the plates at 37°C, overnight.


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	'''Moment of Truth'''		'''Moment of Truth'''

-	'''25.''' Observe the plates under UV-light (312 nm).	+	'''26.''' Observe the plates under UV-light (312 nm).

	'''Note:''' If observed add 254 nm of less intensity of glow		'''Note:''' If observed add 254 nm of less intensity of glow
	could vary.		could vary.
-
-	~~'''Note:''' Steps 17 to 25 explains the protocol for one set~~
-	~~of transformation experiment. Carry out 5 such~~
-	~~transformations simultaneously.~~

File:MG 7177.JPG

2010-09-04T06:40:16Z

uploaded "[[File:MG 7177.JPG]]"

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