Restriction digest - Revision history

Michael A. Speer: /* Specific Protocols */

2009-06-12T20:41:49Z

Specific Protocols

←Older revision		Revision as of 20:41, 12 June 2009
Line 17:		Line 17:
	[[Griffitts:Restriction digest]]		[[Griffitts:Restriction digest]]

-	[[~~Richard_Lab~~:~~Restriction_Digest~~]]	+	[[Richard Lab:Restriction Digest]]

	==Notes==		==Notes==

Michael A. Speer: /* Specific Protocols */

2009-06-12T20:20:27Z

Specific Protocols

←Older revision		Revision as of 20:20, 12 June 2009
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	[[Griffitts:Restriction digest]]		[[Griffitts:Restriction digest]]
		+
		+	[[Richard_Lab:Restriction_Digest]]

	==Notes==		==Notes==

Jason R. Kelly: /* Specific Protocols */

2008-01-28T21:34:18Z

Specific Protocols

←Older revision		Revision as of 21:34, 28 January 2008
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	[[Silver: Restriction Digest]]		[[Silver: Restriction Digest]]
		+
		+	[[Griffitts:Restriction digest]]

	==Notes==		==Notes==

Reshma P. Shetty at 01:27, 9 May 2007

2007-05-09T01:27:18Z

←Older revision		Revision as of 01:27, 9 May 2007
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	*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.		*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.
	*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.		*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.
		+
		+	[[Category:Protocol]]
		+	[[Category:In vitro]]
		+	[[Category:DNA]]

Reshma P. Shetty at 19:31, 8 September 2006

2006-09-08T19:31:51Z

←Older revision		Revision as of 19:31, 8 September 2006
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	*For help with specific enzymes, see [[Restriction enzymes]]		*For help with specific enzymes, see [[Restriction enzymes]]
		+	*For buffer composition, see [[Restriction digest/Buffers]]
	*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.		*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.
	*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.		*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.

Austin J. Che at 00:45, 3 May 2006

2006-05-03T00:45:35Z

←Older revision		Revision as of 00:45, 3 May 2006
Line 17:		Line 17:
	==Notes==		==Notes==

		+	*For help with specific enzymes, see [[Restriction enzymes]]
	*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.		*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.
	*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.		*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.
-	*Improving the efficiency of [[EcoRI/SpeI Double Digest]].
-	*'''Reaction Volumes''' - I used to have very inconsistent results when digesting with EcoRI, using the 20μl reaction volume. Using the Knight Lab's 50μl mix has recently given good results. I haven't tested this enough to say it is a statistically significant result. --[[User:Bcanton\|BC]] 13:39, 2 Jun 2005 (EDT)

Jason R. Kelly: /* Specific Protocols */

2005-09-20T22:03:49Z

Specific Protocols

←Older revision		Revision as of 22:03, 20 September 2005
Line 12:		Line 12:

	[[Knight:Restriction Digest]]		[[Knight:Restriction Digest]]
		+
		+	[[Silver: Restriction Digest]]

	==Notes==		==Notes==

Jason R. Kelly: /* Specific Protocols */

2005-08-25T21:09:10Z

Specific Protocols

←Older revision		Revision as of 21:09, 25 August 2005
Line 7:		Line 7:
	==Specific Protocols==		==Specific Protocols==

-	[[Enzyme ~~Selection~~ for ~~BioBrick Digest~~]]	+	[[Enzyme selection for BioBricks digest]]

	[[Endy:Restriction Digest]]		[[Endy:Restriction Digest]]

Reshma P. Shetty: /* Notes */

2005-07-15T03:37:40Z

Notes

←Older revision		Revision as of 03:37, 15 July 2005
Line 18:		Line 18:
	*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.		*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.
	*Improving the efficiency of [[EcoRI/SpeI Double Digest]].		*Improving the efficiency of [[EcoRI/SpeI Double Digest]].
		+	*'''Reaction Volumes''' - I used to have very inconsistent results when digesting with EcoRI, using the 20μl reaction volume. Using the Knight Lab's 50μl mix has recently given good results. I haven't tested this enough to say it is a statistically significant result. --[[User:Bcanton\|BC]] 13:39, 2 Jun 2005 (EDT)

Reshma P. Shetty at 03:31, 15 July 2005

2005-07-15T03:31:00Z

←Older revision		Revision as of 03:31, 15 July 2005
Line 7:		Line 7:
	==Specific Protocols==		==Specific Protocols==

-	[[Restriction Digest ~~(Biobricks)~~]]	+	[[Enzyme Selection for BioBrick Digest]]
		+
		+	[[Endy:Restriction Digest]]
		+
		+	[[Knight:Restriction Digest]]

	==Notes==		==Notes==
		+
		+	*If you are interested in cutting near the ends of the linear DNA fragment, note that some enzymes do not cut efficiently at the ends of linear DNA. So include extra bases to increase the efficiency of cutting. Many enzymes work with 4 bases supposedly but XhoI was found to require more than 4 bases (8 bases was used successfully). Thus, to be on the safe side, use 8 bases whenever possible. [http://www.neb.com/ NEB] has more information [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_linearized_vector.asp here]. Read the information at NEB carefully ... they recommend adding 4 bases to the numbers listed in their table.
		+	*Tom was once having some trouble with failed restriction digests. He called NEB and they recommended doing the digest in a larger volume. Therefore, the Knight lab typically does digests in either 50 μL or 100 μL volumes rather than the 20 μL volume that the Endy lab uses. If the sample needs to be concentrated, some method of [[Purification of DNA \| DNA purification]] is used.
		+	*Improving the efficiency of [[EcoRI/SpeI Double Digest]].