Bacterial Colony PCR
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Bacterial Colony PCR
Objective:
This protocol allows rapid detection of transformation success when primers are available to allow determination of correct ligation products by size or hybridization.
Procedures:
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Start => Bacterial colonies from transformation.
- Prepare 2mL culture tubes w/ an appropriate selection media for your plasmid of interest, label 1.5mL tubes and place inside culture tube as a cap.
- Prepare PCR master mix: buffer, MgCl2, dNTPs, primers, sucrose red, and enzyme- Aliquot 25µL of master mix into a PCR tube for each colony to be picked. (See general PCR instructions for more instructions on how to set up the PCR reaction.)
- Select colonies to pick. Pick colony with a sterile toothpick or pipet tip. Dab the toothpick or pipet tip into the master mix then place the toothpick or pipet tip in a correspondingly labeled culture tube.
- Run PCR w/ appropriate conditions for your primers and expected product, Making sure to begin your PCR protocol with an extended time at 95°, (e.g. 5 minutes).
- Grow the 2mL cultures O/N at 37°.
- Run 10µL of the PCR reaction on an agarose gel to identify which cultures to keep for plasmid DNA isolation.
PCR方法相关产品:
- 电泳设备
- 紫外设备
- 普通PCR仪
- 定量PCR仪
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PCR/RT-PCR/qPCR试剂
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PCR引物
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PCR试剂
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PCR对照
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特异性PCR试剂盒
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PCR克隆试剂盒
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RNA
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RNase检测/去除
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RT-PCR试剂
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RT-PCR标准品
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定量PCR试剂
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定量PCR标记
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总RNA分离纯化盒
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PCR产物纯化
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核酸酶
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聚合酶
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反转录酶