LDLR Genotyping - Revision history

Davebrid1: Removed table of contents

2026-04-01T12:40:56Z

Removed table of contents

← Older revision		Revision as of 12:40, 1 April 2026
Line 1:		Line 1:
	Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075		Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075

			__NOTOC__

	* [[SOP - Ethidium Bromide]]		* [[SOP - Ethidium Bromide]]

2026-01-26T16:02:31Z

Additional Reagents: Added link to tail lysates

← Older revision		Revision as of 16:02, 26 January 2026
Line 16:		Line 16:
	=== Additional Reagents ===		=== Additional Reagents ===
	* Nuclease-free water		* Nuclease-free water
	* Template DNA (tail lysate or purified genomic DNA)		* Template DNA ([[Preparation of Tail Samples (for Genotyping)\|tail lysate]] or purified genomic DNA)

	== Primer Preparation ==		== Primer Preparation ==

2026-01-26T16:01:47Z

Stock Primer Preparation (100 µM): details about primer resuspension

← Older revision		Revision as of 16:01, 26 January 2026
Line 21:		Line 21:

	=== Stock Primer Preparation (100 µM) ===		=== Stock Primer Preparation (100 µM) ===
	# Resuspend each lyophilized primer in nuclease-free water to 100 µM concentration		# Resuspend each lyophilized primer in nuclease-free water to 100 µM concentration. Look on the tube from IDT, for example if there is 23.7 nmoles then you would resuspend in 237 µL of water
	# Store at -20°C		# Store at -20°C

2026-01-26T16:01:08Z

PCR Primers: Minor typo

← Older revision		Revision as of 16:01, 26 January 2026
Line 7:		Line 7:

	=== PCR Primers ===		=== PCR Primers ===
	* '''Ldlr Common Forward: 5'-TAT GCA TCC CCA GTC TTT GG-3'		* '''Ldlr Common Forward''': 5'-TAT GCA TCC CCA GTC TTT GG-3'
	* '''Ldlr Wild-type Reverse''': 5'-CTA CCC AAC CAG CCC CTT AC-3'		* '''Ldlr Wild-type Reverse''': 5'-CTA CCC AAC CAG CCC CTT AC-3'
	* '''Ldlr Mutant Reverse''': 5'-ATA GAT TCG CCC TTG TGT CC-3'		* '''Ldlr Mutant Reverse''': 5'-ATA GAT TCG CCC TTG TGT CC-3'

2026-01-26T14:53:07Z

PCR Cycling Program

← Older revision		Revision as of 14:53, 26 January 2026
Line 54:		Line 54:

	== PCR Cycling Program ==		== PCR Cycling Program ==

			This program takes 1:50 to run.

	{\| class="wikitable"		{\| class="wikitable"
	\|+ Ldlr ~~Genotyping~~ PCR Program		\|+ '''Ldlr''' PCR Program
	\|-		\|-
	! Step !! Temperature (°C) !! Time !! Cycles !! Notes		! Step !! Temperature (°C) !! Time !! Cycles !! Notes
Line 80:		Line 82:
	\| Final Extension \|\| 72 \|\| 5 min \|\| 1 \|\|		\| Final Extension \|\| 72 \|\| 5 min \|\| 1 \|\|
	\|-		\|-
	\| Hold \|\| 10 \|\| ∞ \|\| \|\|		\| Hold \|\| 4 \|\| ∞ \|\| \|\|
	\|-		\|-
	\|}		\|}

2026-01-26T14:52:31Z

Reagents Needed: Updated primer names

← Older revision		Revision as of 14:52, 26 January 2026
Line 7:		Line 7:

	=== PCR Primers ===		=== PCR Primers ===
	* '''~~Primer 19799''' (~~Common Forward): 5'-TAT GCA TCC CCA GTC TTT GG-3'		* '''Ldlr Common Forward: 5'-TAT GCA TCC CCA GTC TTT GG-3'
	* '''~~Primer 19800~~''' ~~(Wild type Reverse)~~: 5'-CTA CCC AAC CAG CCC CTT AC-3'		* '''Ldlr Wild-type Reverse''': 5'-CTA CCC AAC CAG CCC CTT AC-3'
	* '''~~Primer oIMR7770~~''' ~~(Mutant Reverse)~~: 5'-ATA GAT TCG CCC TTG TGT CC-3'		* '''Ldlr Mutant Reverse''': 5'-ATA GAT TCG CCC TTG TGT CC-3'

	=== Master Mix ===		=== Master Mix ===

2026-01-24T19:38:27Z

formatting

← Older revision		Revision as of 19:38, 24 January 2026
Line 1:		Line 1:
	Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075		Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075

	[[SOP - Ethidium Bromide]]		* [[SOP - Ethidium Bromide]]
	[[SOP - Electrophoresis]]		* [[SOP - Electrophoresis]]

	== Reagents Needed ==		== Reagents Needed ==

2026-01-24T19:38:11Z

Added SOP's

← Older revision		Revision as of 19:38, 24 January 2026
Line 1:		Line 1:
	Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075		Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075

			[[SOP - Ethidium Bromide]]
			[[SOP - Electrophoresis]]

	== Reagents Needed ==		== Reagents Needed ==

2026-01-24T19:33:35Z

Modifieed protocol from Jax, as modified by Claude to use dreamtaq

@@ Line 1: / Line 1: @@
-Based on https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075
+Modified from https://www.jax.org/Protocol?stockNumber=002207&protocolID=27075
-LDLR Genotyping Common - Fwd 	TAT GCA TCC CCA GTC TTT GG
+== Reagents Needed ==
-LDLR Genotyping WT - Rev 	CTA CCC AAC CAG CCC CTT AC
-LDLR Genotyping Mutant  - Rev 	ATA GAT TCG CCC TTG TGT CC
+=== PCR Primers ===
 [[ Category:Genotyping ]]
 [[ Category:Mouse Work ]]
 [[ Category:PCR ]]

2026-01-20T18:11:05Z

Added tags

← Older revision		Revision as of 18:11, 20 January 2026
Line 5:		Line 5:
	LDLR Genotyping Mutant - Rev ATA GAT TCG CCC TTG TGT CC		LDLR Genotyping Mutant - Rev ATA GAT TCG CCC TTG TGT CC

	[[ Genotyping ]]		[[ Category:Genotyping ]]
	[[ Mouse Work ]]		[[ Category:Mouse Work ]]
	[[ PCR ]]		[[ Category:PCR ]]