Ready-to-use multi-probe hybridization slide, and preparation method and application thereof
A multi-probe and probe technology, applied in the field of molecular biology, can solve problems such as time-consuming and laborious, increasing the economic burden of patients, and overnight hybridization
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Embodiment 1F
[0065] The preparation of embodiment 1FISH probe
[0066] This example demonstrates the preparation process of the rapid detection probe for CBFB gene breakage, which specifically includes the following steps:
[0067] (1) Download the genome non-repeated sequence of the region covered by the CBFB gene probe from UCSC Genome Browser, where the CBFB upstream probe coverage area is: chr16:66885331-67101057, and the CBFB downstream probe coverage area is: chr16:67122220-67337946; The obtained sequence is saved in fasta format, and the repeated sequence region in the genome is replaced by N;
[0068] (2) Use the perl plug-in program chunks.pl for the genome non-repeated sequence of the region covered by the CBFB gene upstream probe obtained in step (1) (the chunks.pl program uses the code diagram see Figure 4 ) into blocks of 1kb size; import the divided blocks into OligoArray software in batches for probe design and probe screening. The conditions for probe screening are: probe...
Embodiment 2
[0076] The preparation of embodiment 2 ready-to-use multi-probe hybridization slides
[0077] The present embodiment has invented a kind of preparation method of ready-to-use multi-probe hybridization glass slide
[0078] The probe combinations used in this example were: PML / RARA, BCR / ABL, P53, D13S319, 1Q21, IGH / BCL2, CBFB, AML / ETO; in the corresponding probe attachment area. ( Figure 7 )
[0079] The hybrid slides used in this embodiment are made by entrusting the manufacturer according to the design drawing of the present invention.
[0080] The following is the preparation process of the ready-to-use multi-probe hybridization slide (M-probe Slide):
[0081] (1) According to Figure 1 ~ Figure 3 Probe-attached slides and sample-attached slides prepared by commissioned manufacturers as indicated;
[0082] (2) Carrying out probe attachment in the probe attachment region of the probe attachment glass slide;
[0083] (2.1) According to the formula: levoside 0.5% acacia ...
Embodiment 3
[0088] The method for using the ready-to-use multi-probe hybridization slide comprises the following steps:
[0089] This example demonstrates the usage method and hybridization effect of the ready-to-use multi-probe hybridization slide.
[0090] The probes included in the ready-to-use multi-probe hybridization slide used in this example are: PML / RARA, BCR / ABL, P53, D13S319, 1Q21, IGH / BCL2, CBFB, AML / ETO;
[0091] The sample detected in this embodiment is bone marrow blood lymphocytes fixed by Carnoy's fixative:
[0092] (1) Sample treatment: Add the fixed peripheral blood cells dropwise to the corresponding areas of the sample-attached slides, immerse the sample-attached slides in a container filled with 2×SSC at 37°C for 30 minutes, and pass 70% after the aging is completed. 85%, 100% gradient alcohol dehydration, dry, set aside.
[0093] (2) Probe sample covariance: Add 2ul of hybridization buffer dropwise to the probe attachment area of the hybridization slide, and tur...
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