RT-LAMP对照试剂盒

RT-LAMP对照试剂盒，RT-LAMP Control Kit，用于RT-LAMP检测质量控制的引物和模板试剂盒。更多视频请关注视频号【艾维缔】。哔哩哔哩【IVDSHOW】。抖音【军哥聊表观】。

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RT-LAMP对照试剂盒

所属分类

分子生物学

目录号

PCR544

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For general laboratory use.

Please centrifuge briefly before opening (volume ≤2 ml).

Shipping: shipped on gel packs

Storage Conditions: store at -20 °C
it is highly recommended to aliquot the Total RNA to avoid multiple freezing-thawing cycles

Shelf Life: 12 months

Form: liquid

Description:
RT-LAMP Control Kit contains an optimised mix of 6 primers (F3, B3, FIP, BIP, Loop F, Loop B) and total human RNA as template to set up a quality control assay for RT-LAMP reactions.

Content:

Component	Cap	Conc.	Amount
RT-LAMP Primer Mix	yellow	10x	200 μl
Total RNA - human tissue	red	100 ng/μl	10 μl

Assay setup
Depending on the setup used, a reaction volume of 20-50 μl is recommended for most applications. Pipette with sterile filter tips and perform the setup in a separate area from the DNA preparation or analysis. No-template controls should be included with all amplifications.

The 10x conc. primer mix contains 6 primers in the following concentrations:

included primers	stock conc.
FIP	16 μM
BIP	16 μM
F3	2 μM
B3	2 μM
Loop F	4 μM
Loop B	4 μM

Set up for an isothermal amplification assay
Use the kit in combination with Saphir RT-LAMP Kit (PCR-540). Prepare a master mix as follows:

component	stock conc.	final conc.	20 μl	50 μl
RT-LAMP Buffer (PCR-540)	5 x	1 x	4 μl	10 μl
RT-LAMP Enzymes (PCR-540)	20 x	1 x	1 μl	2.5 μl
RT-LAMP Dye (PCR-540)	25 x	1 x	0.8 μl	2 μl
Primer Mix	10 x	1 x	2 μl	5 μl
Template RNA		<100 ng/assay	x μl	x μl
PCR-grade Water			fill up to 20 μl	fill up to 50 μl

Amplification and detection

Use a specific detection instrument for isothermal amplification or a real-time PCR cycler to run the assays
Set the instrument to 65°C
Select the green channel of the real-time instrument (λexc 494 nm, λem 521 nm)
Measure the fluorescence intensity at an interval of 1 min for up to 30 min.

Trouble shooting
If amplification in no-template controls occurs the following points should be reviewed.

Cross contamination from environments

Clean equipment and areas with "RNA/DNA Away” solution
Replace reagent stocks and pre-mixes with new components
Stop reactions at an earlier point of time before non-template amplification occurs

Carry-over contamination from previous reaction products

Avoid opening reaction vessels after amplification
Use separate preparation area and equipment if post-reaction processing is necessary

Non-template amplification from primers

Increase incubation temperature stepwise by 1-2 °C

产品组分

组分内容	型号	规格	储存温度
RT-LAMP对照试剂盒	PCR544-01	100次	-20°C
RT-LAMP对照试剂盒	PCR544-02	200次	-20°C
操作手册	1	1	室温

*注意：在使用之前，应将溶液离心至管底。

文件资源

营销中心

注意事项

保存建议	推荐蓝冰或冰袋运输。当您收到产品后，按照说明书建议保存各组分。
警告	本品仅供科研使用，请勿用于临床与诊断。

FAQ

1. RT-PCR反应后，无PCR产物或出现Smear，怎么办？
此时请从提取的RNA样品的纯度和添加量、基因的表达丰度、基因的长度和GC含量、引物的设计情况、参考文献的可信度以及RT-PCR条件的设定等方面加以考虑。
2. PCR反应时，反转录反应液的使用量使用多少较为合适？
反转录反应液的使用量可控制在1 μl～10 μl的范围。
3. 3′RACE PCR扩增产物经电泳分析后，有时为什么出现多条带现象？
可能是因为未知序列信息不清楚导致了非特异性扩增；扩增的基因为多基因家族的成员；mRNA不同的拼接方式造成的。