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详细BACDNAIsolation
A. Culture Growth:1.For BAC isolation for shotgun library construction:Pick a sm
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Phenol/ChloroformPrecipitationofDNA
1. Add an equal volume (equal to sample volume) of P/C to sample.2. Mix (shake,
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DNA Purification from Gels
1. Remove gel slice contain DNA fragment and place in 10 volumes of:300 mM NaOAc
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MINI PREP BY BOILING
Pellet 1.5 ml of an overnight culture at 12,000 rpm in Eppendorf centrifuge at R
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Preparation of linear polyacrylamide
Preparation of linear polyacrylamide used as carrier in ethanol precipitation of
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DNAPlasmidPrep.
This is a scaled up version of the Baron protocol which has been modified to ach
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Quick DNA Plasmid Prep
This protocol gives very clean plasmid preps for restriction digests and cloning
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DNAPurificationandPrecipitation
1.Prepare or obtain Buffered phenol,pH 8.0.Add 2 crystals of 8-Hydroxyquinoline
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MITOCHONDRIALDNAISOLATION
1.Grind in mortar and pestle or Waring blender with 5-7 volumes buffer A per g t
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Dialysis (using dialysis tubing)
Molecularpourous membrane tubing is used for desalting, protein and DNA isolatio
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Generation of uni-directionalnesteddeletion sin plasmid DNA
1) Digest DNA with two restriction enzymes such that one end is ExoIII susceptib
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ExtractionfromCheekCellsDNA
Sample PreparationCheek cells are obtained by rinsing the mouth with 25mls of an
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PurificationofPCRfragmentsforcloning
After an aliquot of the PCR mixture is analyzed on an agarose gel, the remainder
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De-phosphorylation of DNA
The preferred method of de-phosphorylation uses the buffer system described by P
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Recoveryof DNA from LMP(Low Melting Point)AgaroseGel
Recovery of DNA from LMP (Low Melting Point) Agarose Gel1. Separate DNA fragment
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