Echinobase/bac information - Revision history

Ctelmer at 20:53, 11 March 2022

2022-03-11T20:53:11Z

← Older revision		Revision as of 20:53, 11 March 2022
Line 54:		Line 54:
	<p style="margin-left: 20px">[[File:Qbot.JPG\|frameless\|200px]]</p>		<p style="margin-left: 20px">[[File:Qbot.JPG\|frameless\|200px]]</p>
	<p>The libraries fit into six -80°C freezers.</p>		<p>The libraries fit into six -80°C freezers.</p>
	<p style="margin-left: 20px">[[File:~~Freezers_hall~~.~~png~~\|frameless\|200px]]</p>		<p style="margin-left: 20px">[[File:Freezer rm.JPG\|frameless\|200px]]</p>
	<p>The truck had power outlets to plug the freezers for the cross-country drive.</p>		<p>The truck had power outlets to plug the freezers for the cross-country drive.</p>
	<p style="margin-left: 20px">[[File:Freezer_truck.png\|frameless\|200px]]</p>		<p style="margin-left: 20px">[[File:Freezer_truck.png\|frameless\|200px]]</p>

Ctelmer at 20:50, 11 March 2022

2022-03-11T20:50:57Z

← Older revision		Revision as of 20:50, 11 March 2022
Line 52:		Line 52:
	<br>'''Transport to Pittsburgh'''		<br>'''Transport to Pittsburgh'''
	<p>The BAC libraries were prepared using the Qbot.</p>		<p>The BAC libraries were prepared using the Qbot.</p>
			<p style="margin-left: 20px">[[File:Qbot.JPG\|frameless\|200px]]</p>
	<p>The libraries fit into six -80°C freezers.</p>		<p>The libraries fit into six -80°C freezers.</p>
			<p style="margin-left: 20px">[[File:Freezers_hall.png\|frameless\|200px]]</p>
	<p>The truck had power outlets to plug the freezers for the cross-country drive.</p>		<p>The truck had power outlets to plug the freezers for the cross-country drive.</p>
			<p style="margin-left: 20px">[[File:Freezer_truck.png\|frameless\|200px]]</p>
	<p>The truck was loaded.</p>		<p>The truck was loaded.</p>
			<p style="margin-left: 20px">[[File:Freezer_truck_inside.png\|frameless\|200px]]</p>
	<p>Ready to go!</p>		<p>Ready to go!</p>
			<p style="margin-left: 20px">[[File:Truck_loaded.png\|frameless\|200px]]</p>
	<p>The CIT Team.</p>		<p>The CIT Team.</p>
			<p style="margin-left: 20px">[[File:CIT group_8956mrgsmall.jpg\|frameless\|200px]]</p>

Ctelmer at 20:42, 11 March 2022

2022-03-11T20:42:27Z

← Older revision		Revision as of 20:42, 11 March 2022
Line 6:		Line 6:
	A subset of the ''S. purpuratus'' BACs have been mapped to the v3.1 genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse on the v5.0 genome assembly. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]).		A subset of the ''S. purpuratus'' BACs have been mapped to the v3.1 genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse on the v5.0 genome assembly. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]).

	'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleagues. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].		<br>'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleagues. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].

	<br>'''Relevant publications:'''		<br>'''Relevant publications:'''
Line 25:		Line 25:
	<p>2. Request the BAC by emailing Veronica Hinman.</p>		<p>2. Request the BAC by emailing Veronica Hinman.</p>

	<p>'''BAC numbers eg. Sp_117A03_L'''</p>		<br><p>'''BAC numbers eg. Sp_117A03_L'''</p>
	<p>Sp = Species Sp, ''S. purpuratus''</p>		<p>Sp = Species Sp, ''S. purpuratus''</p>
	<p>117 = Plate number</p>		<p>117 = Plate number</p>
Line 32:		Line 32:
	<p>L = long or S, short</p>		<p>L = long or S, short</p>

	<br>~~<p>'''Working with BACs'''</p>~~		<br><p>'''Storing BACs'''</p>
	<p>'''Storing BACs'''</p>

	'''Short term'''		'''Short term'''
	to avoid shearing the long BAC DNA fragments, isolated BACs should never be vortexed or frozen. Store BAC DNA at 4°C in TE (10 mM Tris, pH = 7.4; 10 mM EDTA).		<p>To avoid shearing the long BAC DNA fragments, isolated BACs should never be vortexed or frozen. Store BAC DNA at 4°C in TE (10 mM Tris, pH = 7.4; 10 mM EDTA).</p>
	Long term storage
	Store glycerol bacteria stocks (15-25% glycerol) at -80°C.		'''Long term storage'''
			<p>Store glycerol bacteria stocks (15-25% glycerol) at -80°C.</p>

			<br><p>'''Working with BACs'''</p>

	<p>'''Antibiotic conditions''' for working with BAC clones</p>		<p>'''Antibiotic conditions''' for working with BAC clones</p>
Line 47:		Line 49:
	<p>Chloramphenicol Stock (25 μg/μl): Dissolve in ETOH store at -20°C.</p>		<p>Chloramphenicol Stock (25 μg/μl): Dissolve in ETOH store at -20°C.</p>
	<p>Use at 1:2000 dilution (12.5 μg/ml).</p>		<p>Use at 1:2000 dilution (12.5 μg/ml).</p>

			<br>'''Transport to Pittsburgh'''
			<p>The BAC libraries were prepared using the Qbot.</p>

			<p>The libraries fit into six -80°C freezers.</p>

			<p>The truck had power outlets to plug the freezers for the cross-country drive.</p>

			<p>The truck was loaded.</p>

			<p>Ready to go!</p>

			<p>The CIT Team.</p>

Ctelmer at 00:39, 11 March 2022

2022-03-11T00:39:34Z

imported>Ctelmer: /* Genomic BAC Resources */

2021-05-04T19:34:58Z

Genomic BAC Resources

← Older revision		Revision as of 19:34, 4 May 2021
Line 32:		Line 32:
	<p>L = long or S, short</p>		<p>L = long or S, short</p>

	<br>'''Working with BACs'''		<br><p>'''Working with BACs'''</p>
	'''Storing BACs'''		<p>'''Storing BACs'''</p>

	'''Short term'''		'''Short term'''

imported>Ctelmer: /* Genomic BAC Resources */

2021-05-04T19:32:38Z

Genomic BAC Resources

← Older revision		Revision as of 19:32, 4 May 2021
Line 19:		Line 19:

	<br>'''Recombinant BAC Resources'''		<br>'''Recombinant BAC Resources'''
	Echinobase has generated and maintains a set of reporter BAC constructs in which a portion of the gene of interest is replaced by the sequence encoding a fluorescent protein (i.e. GFP, mCherry~~). A list of these sequences is available here~~). Existing clones are available by request. We will also consider requests to make additional recombinant reporter constructs as time permits.		Echinobase has generated and maintains a set of reporter BAC constructs in which a portion of the gene of interest is replaced by the sequence encoding a fluorescent protein (i.e. GFP, mCherry). Existing clones are available by request. We will also consider requests to make additional recombinant reporter constructs as time permits.

	<p>To request a recombinant or wildtype BAC for non-commercial use:</p>		<p>To request a recombinant or wildtype BAC for non-commercial use:</p>
	<p>1. Identify the BAC of interest computationally or by screening our libraries ~~(available filters)~~.</p>		<p>1. Identify the BAC of interest computationally or by screening our libraries.</p>
	<p>2. Request the BAC by emailing Veronica Hinman.</p>		<p>2. Request the BAC by emailing Veronica Hinman.</p>

			<p>'''BAC numbers eg. Sp_117A03_L'''</p>
			<p>Sp = Species Sp, ''S. purpuratus''</p>
			<p>117 = Plate number</p>
			<p>A = Plate row</p>
			<p>03 = Plate column</p>
			<p>L = long or S, short</p>

	'''Working with BACs'''		<br>'''Working with BACs'''
	'''Storing BACs'''		'''Storing BACs'''

Line 34:		Line 40:
	Store glycerol bacteria stocks (15-25% glycerol) at -80 C.		Store glycerol bacteria stocks (15-25% glycerol) at -80 C.

	'''Antibiotic conditions''' for working with BAC clones		<p>'''Antibiotic conditions''' for working with BAC clones</p>
	Kanamycin		<p>Kanamycin Stock (25 μg/μL): Dissolve 0.5 g of kanamycin into 10 ml of ddH2O.</p>
	Stock (25 μg/μL): Dissolve 0.5 g of kanamycin into 10 ml of ddH2O. Filter through a 0.22 µm filter to sterilize. Aliquot and store at -20°C.		<p>Filter through a 0.22 µm filter to sterilize.</p>
	Use at 1:1000 dilution (25 μg/mL).		<p>Aliquot and store at -20°C.</p>
	Chloramphenicol		<p>Use at 1:1000 dilution (25 μg/mL).</p>
	Stock (25 μg/μl): Dissolve in ETOH store at -20.		<p>Chloramphenicol Stock (25 μg/μl): Dissolve in ETOH store at -20.</p>
	Use at 1:2000 dilution.(12.5 μg/ml)		<p>Use at 1:2000 dilution (12.5 μg/ml).</p>

imported>Ctelmer: /* Genomic BAC Resources */

2021-05-04T19:12:46Z

Genomic BAC Resources

← Older revision		Revision as of 19:12, 4 May 2021
Line 8:		Line 8:
	'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleages. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].		'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleages. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].

	'''Relevant publications:'''		<br>'''Relevant publications:'''
	A sea urchin genome project: Sequence scan, virtual map, and additional resources. [http://www.pnas.org/content/97/17/9514.full Cameron, RA, et al., 2000. Proc. Natl. Acad. Sci. USA.]		A sea urchin genome project: Sequence scan, virtual map, and additional resources. [http://www.pnas.org/content/97/17/9514.full Cameron, RA, et al., 2000. Proc. Natl. Acad. Sci. USA.]
	Bacterial artificial chromosomes as recombinant reporter constructs to investigate gene expression and regulation in echinoderms. [http://legacy.echinobase.org/Echinobase/bac_information Buckley, KM et al., 2017. Briefings in Functional Genomics].		Bacterial artificial chromosomes as recombinant reporter constructs to investigate gene expression and regulation in echinoderms. [http://legacy.echinobase.org/Echinobase/bac_information Buckley, KM et al., 2017. Briefings in Functional Genomics].

	[https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library'''BAC Protocols''']		<br><p>[https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library'''BAC Protocols''']</p>
	Screening a BAC library		<p>Screening a BAC library</p>
	Generating a recombinant BAC		<p>Generating a recombinant BAC</p>
	BAC miniprep protocol		<p>BAC miniprep protocol</p>
	Analyzing BAC DNA by PFGE		<p>Analyzing BAC DNA by PFGE</p>

	'''Recombinant BAC Resources'''		<br>'''Recombinant BAC Resources'''
	Echinobase has generated and maintains a set of reporter BAC constructs in which a portion of the gene of interest is replaced by the sequence encoding a fluorescent protein (i.e. GFP, mCherry). A list of these sequences is available here). Existing clones are available by request. We will also consider requests to make additional recombinant reporter constructs as time permits.		Echinobase has generated and maintains a set of reporter BAC constructs in which a portion of the gene of interest is replaced by the sequence encoding a fluorescent protein (i.e. GFP, mCherry). A list of these sequences is available here). Existing clones are available by request. We will also consider requests to make additional recombinant reporter constructs as time permits.

	To request a recombinant or wildtype BAC for non-commercial use:		<p>To request a recombinant or wildtype BAC for non-commercial use:</p>
	1. Identify the BAC of interest computationally or by screening our libraries (available filters).		<p>1. Identify the BAC of interest computationally or by screening our libraries (available filters).</p>
	2. Request the BAC by emailing Veronica Hinman.		<p>2. Request the BAC by emailing Veronica Hinman.</p>

imported>Ctelmer: /* Genomic BAC Resources */

2021-05-04T19:07:11Z

Genomic BAC Resources

← Older revision		Revision as of 19:07, 4 May 2021
Line 4:		Line 4:
	Echinobase stores libraries of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostome species ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]). These libraries were originally constructed as part of the Sea Urchin Genome Project. On average, the insert sizes for our BAC libraries are ~140 kb. Thus, for the 800 Mb ''S. purpuratus'' genome, 1X coverage is encompassed by, on average, 5,700 clones. Each library consists of >100,000 clones, providing ~17X genome coverage.		Echinobase stores libraries of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostome species ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]). These libraries were originally constructed as part of the Sea Urchin Genome Project. On average, the insert sizes for our BAC libraries are ~140 kb. Thus, for the 800 Mb ''S. purpuratus'' genome, 1X coverage is encompassed by, on average, 5,700 clones. Each library consists of >100,000 clones, providing ~17X genome coverage.

	A subset of the ''S. purpuratus'' BACs have been mapped to the genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]))		A subset of the ''S. purpuratus'' BACs have been mapped to the genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]))

	'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleages. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].		'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleages. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].

imported>Ctelmer: Created page with " == Genomic BAC Resources == Echinobase stores libraries of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostom..."

2021-05-04T19:06:24Z

Created page with " == Genomic BAC Resources == Echinobase stores libraries of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostom..."

New page

== Genomic BAC Resources ==

Echinobase stores libraries of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostome species ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]). These libraries were originally constructed as part of the Sea Urchin Genome Project. On average, the insert sizes for our BAC libraries are ~140 kb. Thus, for the 800 Mb ''S. purpuratus'' genome, 1X coverage is encompassed by, on average, 5,700 clones. Each library consists of >100,000 clones, providing ~17X genome coverage.

A subset of the ''S. purpuratus'' BACs have been mapped to the genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]))

'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleages. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].

'''Relevant publications:'''
A sea urchin genome project: Sequence scan, virtual map, and additional resources. [http://www.pnas.org/content/97/17/9514.full Cameron, RA, et al., 2000. Proc. Natl. Acad. Sci. USA.]
Bacterial artificial chromosomes as recombinant reporter constructs to investigate gene expression and regulation in echinoderms. [http://legacy.echinobase.org/Echinobase/bac_information Buckley, KM et al., 2017. Briefings in Functional Genomics].

[https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library'''BAC Protocols''']
Screening a BAC library
Generating a recombinant BAC
BAC miniprep protocol
Analyzing BAC DNA by PFGE

'''Recombinant BAC Resources'''
Echinobase has generated and maintains a set of reporter BAC constructs in which a portion of the gene of interest is replaced by the sequence encoding a fluorescent protein (i.e. GFP, mCherry). A list of these sequences is available here). Existing clones are available by request. We will also consider requests to make additional recombinant reporter constructs as time permits.

To request a recombinant or wildtype BAC for non-commercial use:
1. Identify the BAC of interest computationally or by screening our libraries (available filters).
2. Request the BAC by emailing Veronica Hinman.

'''Working with BACs'''
'''Storing BACs'''

'''Short term'''
to avoid shearing the long BAC DNA fragments, isolated BACs should never be vortexed or frozen. Store BAC DNA at 4 C in TE (10 mM Tris, pH = 7.4; 10 mM EDTA).
Long term storage
Store glycerol bacteria stocks (15-25% glycerol) at -80 C.

'''Antibiotic conditions''' for working with BAC clones
Kanamycin
Stock (25 μg/μL): Dissolve 0.5 g of kanamycin into 10 ml of ddH2O. Filter through a 0.22 µm filter to sterilize. Aliquot and store at -20°C.
Use at 1:1000 dilution (25 μg/mL).
Chloramphenicol
Stock (25 μg/μl): Dissolve in ETOH store at -20.
Use at 1:2000 dilution.(12.5 μg/ml)

← Older revision		Revision as of 00:39, 11 March 2022
Line 2:		Line 2:
	== Genomic BAC Resources ==		== Genomic BAC Resources ==

	~~Echinobase stores libraries~~ of genomic sequence fragments maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostome species ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]). These libraries were originally constructed as part of the Sea Urchin Genome Project. On average, the insert sizes for our BAC libraries are ~140 kb. Thus, for the 800 Mb ''S. purpuratus'' genome, 1X coverage is encompassed by, on average, 5,700 clones. Each library consists of >100,000 clones, providing ~17X genome coverage.		Libraries of genomic sequence fragments are maintained in bacterial artificial chromosome (BAC) vectors for seven lower deuterostome species ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]). These libraries were originally constructed as part of the Sea Urchin Genome Project. On average, the insert sizes for our BAC libraries are ~140 kb. Thus, for the 800 Mb ''S. purpuratus'' genome, 1X coverage is encompassed by, on average, 5,700 clones. Each library consists of >100,000 clones, providing ~17X genome coverage.

	A subset of the ''S. purpuratus'' BACs have been mapped to the genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see ([https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]))		A subset of the ''S. purpuratus'' BACs have been mapped to the v3.1 genome assembly. As part of the SUGP, about 8,000 BAC clones were end sequenced using Sanger technology. Genomic positions for these BACs are indicated as a track in JBrowse on the v5.0 genome assembly. An additional set of individual BACs were sequenced completely; links to the NCBI page are provided in the [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]. Finally, some BACs have been identified using hybridization screening of arrayed library filters (see [https://wiki.echinobase.org/echinowiki/index.php/BAC_Table BAC Table]).

	'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and ~~colleages~~. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].		'''BAC Vector Sequence.''' The BACs are maintained in the pBACe3.6 vector which originates from Children's Hospital Oakland Research Institute in Oakland, California, USA. This vector confers resistance to the antibiotic chloramphenicol and is fully described by Frengen and colleagues. The vector sequence is available [https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library here].

	<br>'''Relevant publications:'''		<br>'''Relevant publications:'''
	A sea urchin genome project: Sequence scan, virtual map, and additional resources. [http://www.pnas.org/content/97/17/9514.full Cameron, RA, et al., 2000. Proc. Natl. Acad. Sci. USA.]		A sea urchin genome project: Sequence scan, virtual map, and additional resources. [http://www.pnas.org/content/97/17/9514.full Cameron, RA, et al., 2000. Proc. Natl. Acad. Sci. USA.]
	Bacterial artificial chromosomes as recombinant reporter constructs to investigate gene expression and regulation in echinoderms. [http://~~legacy~~.echinobase.org/~~Echinobase~~/~~bac_information~~ Buckley, KM et al., 2017. Briefings in Functional Genomics].		Bacterial artificial chromosomes as recombinant reporter constructs to investigate gene expression and regulation in echinoderms. [http://https://www.echinobase.org/literature/article.do?method=display&articleId=45819 Buckley, KM et al., 2017. Briefings in Functional Genomics].

	<br><p>[https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library'''BAC Protocols''']</p>		<br><p>[https://wiki.echinobase.org/echinowiki/index.php/Bac_vector_library'''BAC Protocols''']</p>
Line 36:		Line 36:

	'''Short term'''		'''Short term'''
	to avoid shearing the long BAC DNA fragments, isolated BACs should never be vortexed or frozen. Store BAC DNA at ~~4 C~~ in TE (10 mM Tris, pH = 7.4; 10 mM EDTA).		to avoid shearing the long BAC DNA fragments, isolated BACs should never be vortexed or frozen. Store BAC DNA at 4°C in TE (10 mM Tris, pH = 7.4; 10 mM EDTA).
	Long term storage		Long term storage
	Store glycerol bacteria stocks (15-25% glycerol) at -~~80 C~~.		Store glycerol bacteria stocks (15-25% glycerol) at -80°C.

	<p>'''Antibiotic conditions''' for working with BAC clones</p>		<p>'''Antibiotic conditions''' for working with BAC clones</p>
Line 45:		Line 45:
	<p>Aliquot and store at -20°C.</p>		<p>Aliquot and store at -20°C.</p>
	<p>Use at 1:1000 dilution (25 μg/mL).</p>		<p>Use at 1:1000 dilution (25 μg/mL).</p>
	<p>Chloramphenicol Stock (25 μg/μl): Dissolve in ETOH store at -20.</p>		<p>Chloramphenicol Stock (25 μg/μl): Dissolve in ETOH store at -20°C.</p>
	<p>Use at 1:2000 dilution (12.5 μg/ml).</p>		<p>Use at 1:2000 dilution (12.5 μg/ml).</p>