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DNA extraction from hair...help


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6 replies to this topic

#1 tree

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Posted 02 February 2002 - 08:31 PM

Hi....I need a protocol for extracting DNA from hair/roots...(specifically horse)...and any additional information such as ?storage procedure/timing/etc....thanks....

#2 tree

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Posted 16 February 2002 - 06:37 PM

if anyone is curious, i found a protocol from Quiagen....thanks

#3 diliana

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Posted 23 March 2002 - 06:46 AM

Hi,
Having a look at your problem I remember that I have seen some protocol for DNA extraction from hair in the Bio-Rad site.

Try it and good luck!


#4 Djibrial

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Posted 24 March 2002 - 03:05 AM

I use either of these two methods

1.DNA Extraction from hair:

Cut 10- 15 hair roots about 0.5 cm into a 1.5ml eppendorf tube.
Add 50 ul of 200mM NaOH solution.
Boil the tube in a water bath at 94oC for 10 minutes.
Cool at room temperature and add 50 ul of a solution containing 200mM HCL.
+100mM Tris-HCL pH 8.5.
Your DNA is ready for PCR.
***************************************
2. Another called Dr. Glowatzki's protocol (which seems to work very well for and I prefer to use in most cases):

Cut 10- 15 hair roots about 0.5 cm into a 1.5ml eppendorf tube.
Use 50 ul of the following lysis buffer:
10mM Tris pH 8.3
50mM KCL
0.5% Tween
Also add 10ul of 20ug/ml solution of Proteinase K in 10mM Tris-HCL (pH 7.5)
Vortex for 30 seconds.
Ultracentrifuge at 13000 rpmfor 1 second.
Incubate overnight in a 56 - 60oC waterbath.
Incubate for 10 minutes at 94oC ( to denature the proteinase K  I presume)
Cool down to room temperature.
Ultracentrifuge at 13000 rpm for 1 second.
Ready DNA for your PCR.

Do not hesitate to get in touch incase of any further queries.


#5 Djibrial

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Posted 24 March 2002 - 03:09 AM

I use either of these two methods

1.DNA Extraction from hair:

Cut 10- 15 hair roots about 0.5 cm into a 1.5ml eppendorf tube.
Add 50 ul of 200mM NaOH solution.
Boil the tube in a water bath at 94oC for 10 minutes.
Cool at room temperature and add 50 ul of a solution containing 200mM HCL.
+100mM Tris-HCL pH 8.5.
Your DNA is ready for PCR.
***************************************
2. Another called Dr. Glowatzki's protocol (which seems to work very well for and I prefer to use in most cases):

Cut 10- 15 hair roots about 0.5 cm into a 1.5ml eppendorf tube.
Use 50 ul of the following lysis buffer:
10mM Tris pH 8.3
50mM KCL
0.5% Tween
Also add 10ul of 20ug/ml solution of Proteinase K in 10mM Tris-HCL (pH 7.5)
Vortex for 30 seconds.
Ultracentrifuge at 13000 rpmfor 1 second.
Incubate overnight in a 56 - 60oC waterbath.
Incubate for 10 minutes at 94oC ( to denature the proteinase K  I presume)
Cool down to room temperature.
Ultracentrifuge at 13000 rpm for 1 second.
Ready DNA for your PCR.

Do not hesitate to get in touch incase of any further queries.


#6 tree

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Posted 24 March 2002 - 05:59 AM

Hi,
    I had received more info:  for horse hair, use 2mm of 3 hairs (from the root end) because it is pretty coarse...
    I did this, following the Quiagen protocol, and was able to isolate DNA.....
                        thanks for all the hints....

#7 shaytzur

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Posted 02 March 2005 - 04:44 AM

Hi

Does anyone know how can I purify the hairs before the extraction?
I am sure that the hairs covered with all kinds of bacteria which their DNA will be contain in the final solution.

what is the amount of the DNA extracted from hair?

thanks




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